light-mediated-reversible-modulation-mitogen-activated-protein-kinase

Light-mediated Reversible Modulation of the Mitogen-activated Protein Kinase Pathway during Cell Differentiation and <em>Xenopus</em> Embryonic Development

Kinase activity is crucial for a plethora of cellular functions, including cell proliferation, differentiation, migration, and apoptosis. During early ...

University of Illinois at Urbana-Champaign

The development of skeletal muscle in the vertebrate embryo is controlled by a transcriptional cascade that includes the four myogenic regulatory factors Myf-5, MyoD, Myogenin, and MRF4. The dynamic expression pattern of myf-5 during myogenesis is thought to be consistent with its role during early determination of the myogenic lineage. To study the factors and mechanisms, which regulate myf-5 transcription in Xenopus, we isolated a genomic DNA clone containing 4858 bp of Xmyf-5 5' flanking region. Using a transgenic reporter assay, we show here that this genomic contig is sufficient to recapitulate the dynamic stage- and tissue-specific expression pattern of Xmyf-5 from the gastrula to tail bud stages. For the primary induction of myf-5 transcription, we identify three main regulatory elements, which are responsible for (i) activation in dorsal mesoderm, (ii) activation in ventral mesoderm, and (iii) repression in midline mesoderm, respectively. Their combined activities define the two-winged expression domain of myf-5 in the preinvoluted mesoderm. Repression in midline mesoderm is mediated by a single TCF binding site located in the 5' end of the -4.8 kbp sequence, which binds XTcf-3 protein in vitro. Endogenous Wnt signaling in the lateral mesoderm is required to overcome the long-range repression through this distal TCF site, and to stimulate myf-5 transcription independently from it. The element for ventral mesoderm activation responds to Activin. Together, these results describe a regulatory mosaic of repression and activation, which defines the myf-5 expression profile in the frog gastrula.

Repression through a distal TCF-3 binding site restricts Xenopus myf-5 expression in gastrula mesoderm.

Eye induction and eye field separation are the earliest events during vertebrate eye development. Both of these processes occur much earlier than the formation of optic vesicles. The insulin-like growth factor (IGF) pathway appears to be essential for eye induction, yet it remains unclear how IGF downstream pathways are involved in eye induction. As a consequence of eye induction, a single eye anlage is specified in the anterior neural plate. Subsequently, this single eye anlage is divided into two symmetric eye fields in response to Sonic Hedgehog (Shh) secreted from the prechordal mesoderm. Here, we report that B56epsilon regulatory subunit of protein phosphatase 2A (PP2A) is involved in Xenopus eye induction and subsequent eye field separation. We provide evidence that B56epsilon is required for the IGF/PI3K/Akt pathway and that interfering with the PI3K/Akt pathway inhibits eye induction. In addition, we show that B56epsilon regulates the Hedgehog (Hh) pathway during eye field separation. Thus, B56epsilon is involved in multiple signaling pathways and plays critical roles during early development.

PP2A:B56epsilon is required for eye induction and eye field separation.

Alternative translation is an underappreciated post-transcriptional regulation mechanism. Although only a small number of genes are found to be alternatively translated, most genes undergoing alternative translation play important roles in tumorigenesis and development. Protein phosphatase 2A (PP2A) is involved in many cellular events during tumorigenesis and development. The specificity, localization, and activity of PP2A are regulated by B regulatory subunits. B56epsilon, a member of the B56 regulatory subunit family, is involved in multiple signaling pathways and regulates a number of developmental processes. Here we report that B56epsilon is alternatively translated, leading to the production of a full-length form and a shorter isoform that lacks the N-terminal 76 amino acid residues of the full-length form. Alternative translation of B56epsilon occurs through a cap-dependent mechanism. We provide evidence that the shorter isoform is required for Wnt signaling and regulates the midbrain/hindbrain boundary formation during Xenopus embryonic development. This demonstrates that the shorter isoform of B56epsilon has important biological functions. Furthermore, we show that the N-terminal sequence of B56epsilon, which is not present in the shorter isoform, contains a nuclear localization signal, whereas the C terminus of B56epsilon contains a nuclear export signal. The shorter isoform, which lacks the N-terminal nuclear localization signal, is restricted to the cytoplasm. In contrast, the full-length form can be localized to the nucleus in a cell type-specific manner. The finding that B56epsilon is alternatively translated adds a new level of regulation to PP2A holoenzymes.

The 48-kDa alternative translation isoform of PP2A:B56epsilon is required for Wnt signaling during midbrain-hindbrain boundary formation.

As the key component of many hemoproteins (heme-containing proteins), heme is involved in a broad range of biological processes. Enzymes required for heme biosynthesis and degradation pathways are evolutionarily conserved. While heme metabolism has been studied extensively, the expression of heme metabolism enzymes during development has not been described. Here, we report that all heme biosynthases and two heme oxygenases, which initiate heme degradation, are dynamically expressed during Xenopus embryonic development. All heme synthases, with the exception of aminolevulinic acid synthase 2, are maternally expressed. At neurula stage, heme synthases are expressed in the developing neural tissue and in migrating neural crest cells. At the swimming tadpole stage, expression of heme synthases can be detected in multiple lineages, including eyes, neural crest cells, developing central nervous system, ventral blood island, pronephron, and pronephric tubule. Similar to heme synthases, heme oxygenases are expressed maternally. Zygotic expression of heme oxygenases is mainly restricted to the developing neural and neural crest lineages. Unlike heme synthases, heme oxygenases are not expressed in the ventral blood island and are expressed at a very low level in the pronephron and pronephric tubule. This indicates that heme metabolism may play important roles during development.

Heme metabolism enzymes are dynamically expressed during Xenopus embryonic development.

Regulated intestinal stem cell proliferation and differentiation are required for normal intestinal homeostasis and repair after injury. The Notch signaling pathway plays fundamental roles in the intestinal epithelium. Despite the fact that Notch signaling maintains intestinal stem cells in a proliferative state and promotes absorptive cell differentiation in most species, it remains largely unclear how Notch signaling itself is precisely controlled during intestinal homeostasis. We characterized the intestinal phenotypes of brom bones, a zebrafish mutant carrying a nonsense mutation in hnRNP I. We found that the brom bones mutant displays a number of intestinal defects, including compromised secretory goblet cell differentiation, hyperproliferation, and enhanced apoptosis. These phenotypes are accompanied by a markedly elevated Notch signaling activity in the intestinal epithelium. When overexpressed, hnRNP I destabilizes the Notch intracellular domain (NICD) and inhibits Notch signaling. This activity of hnRNP I is conserved from zebrafish to human. In addition, our biochemistry experiments demonstrate that the effect of hnRNP I on NICD turnover requires the C-terminal portion of the RAM domain of NICD. Our results demonstrate that hnRNP I is an evolutionarily conserved Notch inhibitor and plays an essential role in intestinal homeostasis.

hnRNP I inhibits Notch signaling and regulates intestinal epithelial homeostasis in the zebrafish.

&#x86CB;&#x6FC0;&#x6D3B;&#x662F;&#x9632;&#x6B62;&#x591A;&#x7CBE;&#x53D7;&#x7CBE;&#x548C;&#x53D7;&#x7CBE;&#x5375;&#x7684;&#x542F;&#x52A8;&#x53D1;&#x5C55;&#x6240;&#x9700;&#x7684;&#x91CD;&#x8981;&#x7EC6;&#x80DE;&#x4E8B;&#x4EF6;&#x3002;&#x5BA1;&#x67E5;&#x6240;&#x6709;&#x52A8;&#x7269;&#x7684;&#x86CB;&#x6FC0;&#x6D3B;&#x662F;&#x5728;&#x53D7;&#x7CBE;&#x7684;&#x5375;&#x5B50;&#x7EC6;&#x80DE;&#x8D28;&#x6DB2;&#x4E2D;&#x81EA;&#x7531;&#x7ECF;&#x4E0A;&#x5347;&#x5F15;&#x8D77;&#x7684;&#x3002;&#x6B64;&#x7ECF;&#x4E0A;&#x5347;&#x5BF9;&#x4E8E;&#x6240;&#x6709;&#x7684;&#x540E;&#x7EED;&#x86CB;&#x6FC0;&#x6D3B;&#x6B65;&#x9AA4;&#xFF0C;&#x4F8B;&#x5982;&#x76AE;&#x8D28;&#x9897;&#x7C92;&#x80DE;&#xFF0C;&#x4FEE;&#x6539;&#x5375;&#x9EC4;&#x819C;&#x9632;&#x6B62;&#x591A;&#x7CBE;&#x53D7;&#x7CBE;&#x81F3;&#x5173;&#x91CD;&#x8981;&#x3002;&#x6E38;&#x79BB;&#x7ECF;&#x4E0A;&#x5347;&#x4E3B;&#x8981;&#x7531; 1,4,5-&#x808C;&#x9187; (IP(3))-&#x4ECB;&#x5BFC;&#x5185;&#x8D28;&#x7F51;&#x7ECF;&#x91CA;&#x653E;&#x3002;&#x57FA;&#x56E0;&#x7684;&#x8C03;&#x8282; (3) IP-&#x4ECB;&#x5BFC;&#x7684;&#x7ECF;&#x91CA;&#x653E;&#x86CB;&#x6FC0;&#x6D3B;&#x671F;&#x95F4;&#x4ECD;&#x7136;&#x5F88;&#x5927;&#x7A0B;&#x5EA6;&#x4E0A;&#x672A;&#x77E5;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x62A5;&#x544A;&#x5BF9;&#x6591;&#x9A6C;&#x9C7C;&#x7684;&#x4EA7;&#x5987;&#x5F71;&#x54CD;&#x7A81;&#x53D8;&#x79CD;&#xFF0C;brom &#x9AA8;&#x5934;&#xFF0C;&#x8FD9;&#x662F;&#x5728;&#x6E38;&#x79BB;&#x7ECF;&#x4E0A;&#x5347;&#x5B58;&#x5728;&#x7F3A;&#x9677;&#x548C;&#x968F;&#x540E;&#x86CB;&#x6FC0;&#x6D3B;&#x4E8B;&#x4EF6;&#xFF0C;&#x5176;&#x4E2D;&#x5305;&#x62EC;&#x76AE;&#x8D28;&#x9897;&#x7C92;&#x80DE;&#x548C;&#x7EC6;&#x80DE;&#x8D28;&#x9694;&#x79BB;&#x3002;&#x6211;&#x4EEC;&#x663E;&#x793A;&#x901A;&#x8FC7;&#x63D0;&#x4F9B;&#x7ECF;&#x6216; Ca(2+) &#x91CA;&#x653E;&#x4FE1;&#x4F7F; IP(3)&#xFF0C;&#x8868;&#x660E; brom &#x9AA8;&#x9ABC;&#x7684; IP (3) &#x8C03;&#x8282;&#x5668;&#x5C31;&#x53EF;&#x4EE5;&#x6551;&#x51FA; brom &#x9AA8;&#x5934;&#x7684;&#x86CB;&#x6FC0;&#x6D3B;&#x7F3A;&#x9677;-&#x4ECB;&#x5BFC;&#x7ECF;&#x91CA;&#x653E;&#x5728;&#x65BD;&#x80A5;&#x3002;&#x6709;&#x8DA3;&#x7684;&#x662F;&#xFF0C;brom &#x9AA8;&#x9ABC;&#x7A81;&#x53D8;&#x4F53;&#x80DA;&#x80CE;&#x4E5F;&#x663E;&#x793A;&#x7F3A;&#x9677;&#x4EB2;&#x8F74;&#x5F62;&#x6210;&#x4F34;&#x968F;&#x7740;&#x6742;&#x4E71;&#x65E0;&#x7AE0;&#x7684;&#x76AE;&#x5C42;&#x5FAE;&#x7BA1;&#x7F51;&#x7EDC;&#xFF0C;&#x540E;&#x8005;&#x88AB;&#x8BA4;&#x4E3A;&#x662F;&#x80CC;&#x8F74;&#x5F62;&#x6210;&#x7684;&#x5173;&#x952E;&#x3002;&#x6211;&#x4EEC;&#x63D0;&#x4F9B;&#x53D7;&#x635F;&#x5FAE;&#x7BA1;&#x7EC4;&#x7EC7;&#x662F;&#x4E0E;&#x975E; exocytosed &#x76AE;&#x8D28;&#x9897;&#x7C92;&#x4ECE;&#x8F83;&#x65E9;&#x7684;&#x86CB;&#x6FC0;&#x6D3B;&#x7F3A;&#x9677;&#x5173;&#x8054;&#x7684;&#x8BC1;&#x636E;&#x3002;Brom &#x9AA8;&#x9ABC;&#x57FA;&#x56E0;&#x7684;&#x5B9A;&#x4F4D;&#x514B;&#x9686;&#x663E;&#x793A;&#xFF0C;&#x8FC7;&#x65E9;&#x505C;&#x6B62; hnRNP &#x7684;&#x57FA;&#x56E0;&#x5BC6;&#x7801;&#x5B50;&#x6211; &#xFF08;&#x8FD9;&#x91CC;&#x79F0;&#x4E3A; hnrnp &#x6211;&#xFF09; &#x662F;&#x5F02;&#x5E38;&#x7684;&#x57FA;&#x7840;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x7814;&#x7A76;&#x56E0;&#x6B64;&#x63ED;&#x793A;&#x91CD;&#x8981;&#x7684;&#x65B0;&#x89D2;&#x8272;&#x7684; hnrnp &#x6211;&#x5728;&#x89C4;&#x7BA1;&#x7684; IP (3) &#x57FA;&#x672C;&#x8FC7;&#x7A0B;-&#x4ECB;&#x5BFC;&#x7684;&#x86CB;&#x6FC0;&#x6D3B;&#x7ECF;&#x91CA;&#x653E;&#x3002;

&#x6211;&#x9700;&#x8981;&#x751F;&#x6210;&#x539F;&#x56E0;&#x86CB;&#x5728;&#x6591;&#x9A6C;&#x9C7C;&#x4E2D;&#x7684;&#x6FC0;&#x6D3B;&#x7684;&#x4FE1;&#x53F7; Ca2 + hnRNP&#x3002;

Egg activation is an important cellular event required to prevent polyspermy and initiate development of the zygote. Egg activation in all animals examined is elicited by a rise in free Ca(2+) in the egg cytosol at fertilization. This Ca(2+) rise is crucial for all subsequent egg activation steps, such as cortical granule exocytosis, which modifies the vitelline membrane to prevent polyspermy. The cytosolic Ca(2+) rise is primarily initiated by inositol 1,4,5-trisphosphate (IP(3))-mediated Ca(2+) release from the endoplasmic reticulum. The genes involved in regulating the IP(3)-mediated Ca(2+) release during egg activation remain largely unknown. Here we report on a zebrafish maternal-effect mutant, brom bones, which is defective in the cytosolic Ca(2+) rise and subsequent egg activation events, including cortical granule exocytosis and cytoplasmic segregation. We show that the egg activation defects in brom bones can be rescued by providing Ca(2+) or the Ca(2+)-release messenger IP(3), suggesting that brom bones is a regulator of IP(3)-mediated Ca(2+) release at fertilization. Interestingly, brom bones mutant embryos also display defects in dorsoventral axis formation accompanied by a disorganized cortical microtubule network, which is known to be crucial for dorsal axis formation. We provide evidence that the impaired microtubule organization is associated with non-exocytosed cortical granules from the earlier egg activation defect. Positional cloning of the brom bones gene reveals that a premature stop codon in the gene encoding hnRNP I (referred to here as hnrnp I) underlies the abnormalities. Our studies therefore reveal an important new role of hnrnp I in regulating the fundamental process of IP(3)-mediated Ca(2+) release at egg activation.

hnRNP I is required to generate the Ca2+ signal that causes egg activation in zebrafish.

Activation Egg est un &#xE9;v&#xE9;nement important cellulaire n&#xE9;cessaire pour emp&#xEA;cher la polyspermie et amorcer le d&#xE9;veloppement du zygote. L&#x26;#39;activation d&#x26;#39;oeufs dans tous les animaux examin&#xE9;s est provoqu&#xE9;e par une augmentation de Ca libre (2 +) dans le cytosol oeuf lors de la f&#xE9;condation. Ce Ca (2 +) hausse est crucial pour toutes les &#xE9;tapes ult&#xE9;rieures d&#x26;#39;activation d&#x26;#39;&#x153;ufs, comme exocytose des granules corticaux, qui modifie la membrane vitelline pour emp&#xEA;cher la polyspermie. Le Ca cytosolique (2 +) hausse est principalement initi&#xE9;e par l&#x26;#39;inositol 1,4,5-triphosphate (IP (3)) &#xE0; m&#xE9;diation Ca (2 +) communiqu&#xE9; par le r&#xE9;ticulum endoplasmique. Les g&#xE8;nes impliqu&#xE9;s dans la r&#xE9;gulation de la propri&#xE9;t&#xE9; intellectuelle (3)-m&#xE9;di&#xE9;e Ca (2 +) communiqu&#xE9; lors de l&#x26;#39;activation d&#x26;#39;&#x153;ufs restent encore largement inconnus. Nous rapportons ici un poisson z&#xE8;bre maternelle &#xE0; effet mutant, Brom Bones, qui est d&#xE9;fectueux dans le Ca cytosolique (2 +) hausse et &#xE9;v&#xE9;nements ult&#xE9;rieurs d&#x26;#39;activation d&#x26;#39;&#x153;ufs, y compris exocytose des granules corticaux et la s&#xE9;gr&#xE9;gation cytoplasmique. Nous montrons que les d&#xE9;fauts d&#x26;#39;activation d&#x26;#39;&#x153;ufs dans Brom Bones peut &#xEA;tre sauv&#xE9; en fournissant Ca (2 +) ou le Ca (2 +) IP messager de lib&#xE9;ration (3), ce qui sugg&#xE8;re que brom os est un r&#xE9;gulateur de la propri&#xE9;t&#xE9; intellectuelle (3)-m&#xE9;di&#xE9;e Ca (2 +) communiqu&#xE9; lors de la f&#xE9;condation. Fait int&#xE9;ressant, brom embryons mutants os &#xE9;galement afficher des d&#xE9;fauts dans la formation axe dorso-ventral accompagn&#xE9;e par un r&#xE9;seau de microtubules corticaux d&#xE9;sorganis&#xE9;, ce qui est connu pour &#xEA;tre cruciale pour la formation de l&#x26;#39;axe dorsal. Nous fournir la preuve que l&#x26;#39;organisation des microtubules avec facult&#xE9;s affaiblies est associ&#xE9; &#xE0; la non-exocytosed granules corticaux de l&#x26;#39;anomalie d&#x26;#39;activation oeuf plus t&#xF4;t. Le clonage positionnel du g&#xE8;ne brom os r&#xE9;v&#xE8;le que un codon stop pr&#xE9;matur&#xE9; dans le g&#xE8;ne codant pour hnRNP I (appel&#xE9;e ici hnRNP I) sous-tend les anomalies. Nos &#xE9;tudes r&#xE9;v&#xE8;lent donc un nouveau r&#xF4;le important de hnRNP I dans la r&#xE9;gulation du processus fondamental de la propri&#xE9;t&#xE9; intellectuelle (3)-m&#xE9;di&#xE9;e Ca (2 +) communiqu&#xE9; lors de l&#x26;#39;activation d&#x26;#39;oeuf.

HnRNP I est requis pour g&#xE9;n&#xE9;rer le signal Ca2 + Cela provoque une activation des oeufs chez le poisson z&#xE8;bre

Ei-Aktivierung ist ein wichtiges zellul&auml;re Ereignis, erforderlich um Polyspermie und initiieren Entwicklung der Zygote zu verhindern. Ei-Aktivierung bei allen untersuchten Tieren ist durch einen Anstieg der freien Ca(2+) in der Ei-Zytosol bei Befruchtung l&ouml;ste. Dieser Anstieg des Ca(2+) ist entscheidend f&uuml;r alle nachfolgenden Ei Aktivierungsschritte, wie kortikale Untermodul Exozytose, wodurch die Dotter Membrane um zu verhindern, dass Polyspermie ge&auml;ndert. Der Zytosolische Ca(2+) Aufstieg wird in erster Linie durch Inosit 1,4,5-Trisphosphate (IP(3)) initiiert-vermittelte Ca(2+) Entlassung aus dem Endoplasmatischen Retikulum. Den Genen, die bei der Regulierung der IP (3)-vermittelte Ca(2+) Freigabe w&auml;hrend Ei Aktivierung bleiben weitgehend unbekannt. Hier berichten wir &uuml;ber ein Zebrafisch-m&uuml;tterliche-Effekt-Mutant, Ei Brom Knochen, die Defekte in der zellul&auml;ren Ca(2+) steigen und nachfolgende Aktivierung Veranstaltungen, einschlie&szlig;lich der kortikalen Untermodul Exozytose und zytoplasmatischen Segregation. Wir zeigen, dass die Ei-Aktivierung-Defekte in Brom Knochen gerettet werden k&ouml;nnen, durch die Bereitstellung von Ca(2+) oder die Ca(2+)-Version Messenger IP(3), was darauf hindeutet, dass Brom Knochen ist ein Regulator der IP (3)-vermittelte Ca(2+) Release auf Befruchtung. Interessanterweise zeigen Brom Knochen mutant Embryonen auch M&auml;ngel in dorsoventral Achse-Formation, begleitet von einer desorganisierten kortikale Microtubule-Netzwerk, das entscheidend f&uuml;r dorsale Achse Bildung bekannt ist. Wir belegen, dass die gest&ouml;rte Microtubule-Organisation nicht exocytosed kortikale Granulat aus der fr&uuml;heren Ei Aktivierung Fehler zugeordnet ist. Positionsklonen Brom Knochen gen ergibt, dass ein vorzeitiges Stopp-Codon im Gen f&uuml;r HnRNP ich (genannt hier Hnrnp I) zugrunde liegt die Anomalien. Unsere Studien zeigen daher eine wichtige neue Rolle der Hnrnp ich bei der Regulierung des grundlegenden Prozess der IP (3)-vermittelte Ca(2+) Release auf Ei-Aktivierung.

HnRNP ich erforderlich ist, um das Ca2 +-Signal zu erzeugen, dass Ursachen Aktivierung im Zebrafisch Ei.

Attivazione uovo &egrave; un importante evento cellulare necessario per evitare polyspermy e avviare lo sviluppo dello zigote. Attivazione uovo in tutti gli animali esaminati &egrave; provocata da un aumento della Ca(2+) libero nel citosol uovo alla fecondazione. Questo aumento di Ca(2+) &egrave; cruciale per tutti i passaggi di attivazione uovo successive, ad esempio esocitosi granulo corticale, che modifica la membrana vitellina per evitare polyspermy. L&#39;aumento di Ca(2+) citosolico principalmente &egrave; iniziata da inositolo 1.4.5-trifosfato (IP(3))-mediata Ca(2+) rilascio dal reticolo endoplasmatico. I geni coinvolti nella regolazione dell&#39;IP (3)-mediata Ca(2+) rilascio durante l&#39;attivazione uovo rimangono in gran parte sconosciuto. Qui riportiamo su un mutante di zebrafish effetto materno, brom bones, che &egrave; difettoso in aumento Ca(2+) citosolico e successive uovo eventi di attivazione, compresi esocitosi dei granuli corticali e segregazione citoplasmatica. Mostriamo che i difetti di attivazione uovo di brom bones possono essere salvati fornendo Ca(2+) o il Ca(2+)-release messenger IP(3), suggerendo che brom bones &egrave; un regolatore di IP (3)-mediata Ca(2+) rilascio alla fecondazione. &Egrave; interessante notare che, embrioni mutanti brom bones visualizzare anche difetti nella formazione asse dorsoventral accompagnata da una rete dei microtubuli corticali disorganizzata, che &egrave; noto per essere cruciale per la formazione di asse dorsale. Noi fornire la prova che l&#39;organizzazione alterata dei microtubuli &egrave; associato con granuli corticali non exocytosed dal difetto di attivazione uovo precedente. Clonazione posizionale del gene brom bones rivela che un codone di stop prematuro nel gene che codifica hnRNP ho (denominate hnrnp io) &egrave; alla base delle anomalie. I nostri studi rivelano pertanto un importante ruolo nuovo di hnrnp in che regolano il processo fondamentale di IP (3)-mediata Ca(2+) comunicato all&#39;attivazione di uovo.

hnRNP &egrave; necessaria per generare il segnale Ca2 + che cause uovo attivazione nello zebrafish.

&#x5375;&#x306E;&#x6D3B;&#x6027;&#x5316;&#x306F;&#x3001;&#x591A;&#x7CBE;&#x5B50;&#x53D7;&#x7CBE;&#x3092;&#x9632;&#x6B62;&#x3057;&#x3001;&#x53D7;&#x7CBE;&#x5375;&#x306E;&#x958B;&#x767A;&#x3092;&#x958B;&#x59CB;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x5FC5;&#x8981;&#x306A;&#x91CD;&#x8981;&#x306A;&#x7D30;&#x80DE;&#x306E;&#x30A4;&#x30D9;&#x30F3;&#x30C8;&#x3067;&#x3059;&#x3002;&#x8ABF;&#x3079;&#x305F;&#x5168;&#x3066;&#x306E;&#x52D5;&#x7269;&#x306E;&#x5375;&#x306E;&#x6D3B;&#x6027;&#x5316;&#x306F;&#x53D7;&#x7CBE;&#x306B;&#x304A;&#x3051;&#x308B;&#x5375;&#x7D30;&#x80DE;&#x8CEA;&#x5185;&#x306E;&#x904A;&#x96E2;Ca&#x306E;&#x4E0A;&#x6607;&#xFF08;2 +&#xFF09;&#x306B;&#x3088;&#x3063;&#x3066;&#x8A98;&#x767A;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;Ca&#xFF08;2 +&#xFF09;&#x306E;&#x4E0A;&#x6607;&#x306F;&#x3001;&#x3053;&#x306E;&#x3088;&#x3046;&#x306A;&#x591A;&#x7CBE;&#x3092;&#x9632;&#x3050;&#x305F;&#x3081;&#x5375;&#x9EC4;&#x819C;&#x3092;&#x5909;&#x66F4;&#x3059;&#x308B;&#x76AE;&#x8CEA;&#x306E;&#x9846;&#x7C92;&#x30A8;&#x30AD;&#x30BD;&#x30B5;&#x30A4;&#x30C8;&#x30FC;&#x30B7;&#x30B9;&#x306A;&#x3069;&#x306E;&#x5F8C;&#x7D9A;&#x306E;&#x3059;&#x3079;&#x3066;&#x306E;&#x5375;&#x6D3B;&#x6027;&#x5316;&#x306E;&#x30B9;&#x30C6;&#x30C3;&#x30D7;&#x306E;&#x305F;&#x3081;&#x306B;&#x91CD;&#x8981;&#x3067;&#x3042;&#x308B;&#x3002;&#x7D30;&#x80DE;&#x5185;Ca&#xFF08;2 +&#xFF09;&#x306E;&#x4E0A;&#x6607;&#x306F;&#x3001;&#x4E3B;&#x306B;&#x30A4;&#x30CE;&#x30B7;&#x30C8;&#x30FC;&#x30EB;1,4,5  - &#x4E09;&#x30EA;&#x30F3;&#x9178;&#xFF08;IP 3&#xFF09;&#x3092;&#x4ECB;&#x3057;&#x305F;&#x5C0F;&#x80DE;&#x4F53;&#x304B;&#x3089;&#x306E;Ca&#xFF08;2 +&#xFF09;&#x653E;&#x51FA;&#x306B;&#x3088;&#x3063;&#x3066;&#x958B;&#x59CB;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x5375;&#x6D3B;&#x6027;&#x5316;&#x6642;&#x306B;IP&#xFF08;3&#xFF09;&#x5A92;&#x4ECB;&#x6027;&#x306E;Ca&#xFF08;2 +&#xFF09;&#x653E;&#x51FA;&#x306E;&#x8ABF;&#x7BC0;&#x306B;&#x95A2;&#x4E0E;&#x3059;&#x308B;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x5927;&#x90E8;&#x5206;&#x306F;&#x4E0D;&#x660E;&#x306E;&#x307E;&#x307E;&#x3067;&#x3042;&#x308B;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;&#x7D30;&#x80DE;&#x5185;Ca&#xFF08;2 +&#xFF09;&#x306E;&#x4E0A;&#x6607;&#x3068;&#x76AE;&#x8CEA;&#x9846;&#x7C92;&#x30A8;&#x30AD;&#x30BD;&#x30B5;&#x30A4;&#x30C8;&#x30FC;&#x30B7;&#x30B9;&#x3068;&#x7D30;&#x80DE;&#x8CEA;&#x306E;&#x5206;&#x96E2;&#x3092;&#x542B;&#x3081;&#x3001;&#x305D;&#x306E;&#x5F8C;&#x306E;&#x5375;&#x6D3B;&#x6027;&#x5316;&#x30A4;&#x30D9;&#x30F3;&#x30C8;&#x306B;&#x6B20;&#x9665;&#x304C;&#x3042;&#x308B;&#x30BC;&#x30D6;&#x30E9;&#x30D5;&#x30A3;&#x30C3;&#x30B7;&#x30E5;&#x6BCD;&#x6027;&#x52B9;&#x679C;&#x7A81;&#x7136;&#x5909;&#x7570;&#x4F53;&#x3001;&#x30D6;&#x30ED;&#x30E0;&#x9AA8;&#x306B;&#x3064;&#x3044;&#x3066;&#x5831;&#x544A;&#x3059;&#x308B;&#x3002;&#x6211;&#x3005;&#x306F;&#x30D6;&#x30ED;&#x30E0;&#x9AA8;&#x304C;IP&#x306E;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30BF;&#xFF08;3&#xFF09;&#x5A92;&#x4ECB;&#x6027;&#x306E;Ca&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x5506;&#x3057;&#x3001;&#x30D6;&#x30ED;&#x30E0;&#x306E;&#x9AA8;&#x3067;&#x5375;&#x6D3B;&#x6027;&#x5316;&#x306E;&#x6B20;&#x9665;&#x306F;&#x3001;Ca&#xFF08;2 +&#xFF09;&#x307E;&#x305F;&#x306F;Ca&#xFF08;2 +&#xFF09;&#x653E;&#x51FA;&#x30E1;&#x30C3;&#x30BB;&#x30F3;&#x30B8;&#x30E3;&#x30FC;IP&#xFF08;3&#xFF09;&#x3092;&#x63D0;&#x4F9B;&#x3059;&#x308B;&#x3053;&#x3068;&#x306B;&#x3088;&#x308A;&#x3001;&#x6551;&#x52A9;&#x3067;&#x304D;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x308B;&#xFF08;2 +&#xFF09;&#x53D7;&#x7CBE;&#x3067;&#x30EA;&#x30EA;&#x30FC;&#x30B9;&#x3002;&#x8208;&#x5473;&#x6DF1;&#x3044;&#x3053;&#x3068;&#x306B;&#x3001;&#x30D6;&#x30ED;&#x30E0;&#x9AA8;&#x5909;&#x7570;&#x4F53;&#x306E;&#x80DA;&#x306B;&#x3082;&#x80CC;&#x8EF8;&#x5F62;&#x6210;&#x306B;&#x91CD;&#x8981;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x304C;&#x77E5;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x7121;&#x79E9;&#x5E8F;&#x306A;&#x76AE;&#x8CEA;&#x5FAE;&#x5C0F;&#x7BA1;&#x30CD;&#x30C3;&#x30C8;&#x30EF;&#x30FC;&#x30AF;&#x3092;&#x4F34;&#x3046;&#x80CC;&#x8179;&#x8EF8;&#x306E;&#x5F62;&#x6210;&#x306B;&#x6B20;&#x9665;&#x304C;&#x8868;&#x793A;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x79C1;&#x305F;&#x3061;&#x306F;&#x3001;&#x969C;&#x5BB3;&#x306E;&#x5FAE;&#x5C0F;&#x7BA1;&#x306E;&#x7D44;&#x7E54;&#x306F;&#x3001;&#x4EE5;&#x524D;&#x306E;&#x5375;&#x6D3B;&#x6027;&#x5316;&#x306E;&#x6B20;&#x9665;&#x304B;&#x3089;&#x975E;exocytosed&#x76AE;&#x8CEA;&#x9846;&#x7C92;&#x306B;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x3053;&#x3068;&#x3092;&#x8A3C;&#x62E0;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x30D6;&#x30ED;&#x30E0;&#x9AA8;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x30DD;&#x30B8;&#x30B7;&#x30E7;&#x30CA;&#x30EB;&#x30AF;&#x30ED;&#x30FC;&#x30CB;&#x30F3;&#x30B0;&#x306F;&#x3001;&#x30B3;&#x30FC;&#x30C9;&#x3059;&#x308B;&#x907A;&#x4F1D;&#x5B50;&#x306E;hnRNP I&#xFF08;&#x306E;hnRNP&#x79C1;&#x3068;&#x3057;&#x3066;&#x3053;&#x3053;&#x306B;&#x547C;&#x3070;&#x308C;&#x308B;&#xFF09;&#x306E;&#x57FA;&#x790E;&#x3068;&#x306A;&#x308B;&#x7570;&#x5E38;&#x306E;&#x672A;&#x719F;&#x7D42;&#x6B62;&#x30B3;&#x30C9;&#x30F3;&#x3053;&#x3068;&#x304C;&#x660E;&#x3089;&#x304B;&#x306B;&#x306A;&#x3063;&#x305F;&#x3002;&#x6211;&#x3005;&#x306E;&#x7814;&#x7A76;&#x306F;&#x3001;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x3001;IP&#x306E;&#x57FA;&#x672C;&#x7684;&#x306A;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#xFF08;3&#xFF09;&#x5A92;&#x4ECB;&#x5375;&#x6D3B;&#x6027;&#x5316;&#x6642;&#x306E;Ca&#xFF08;2 +&#xFF09;&#x653E;&#x51FA;&#x3092;&#x8ABF;&#x7BC0;&#x3059;&#x308B;&#x306E;hnRNP&#x79C1;&#x306E;&#x65B0;&#x3057;&#x3044;&#x91CD;&#x8981;&#x306A;&#x5F79;&#x5272;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x305F;&#x3002;

&#x306E;hnRNP I&#x306F;&#x3001;&#x30BC;&#x30D6;&#x30E9;&#x30D5;&#x30A3;&#x30C3;&#x30B7;&#x30E5;&#x306E;&#x5375;&#x6D3B;&#x6027;&#x5316;&#x3092;&#x5F15;&#x304D;&#x8D77;&#x3053;&#x3059;Ca2 +&#x306E;&#x4FE1;&#x53F7;&#x3092;&#x751F;&#x6210;&#x3059;&#x308B;&#x5FC5;&#x8981;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;

&#xACC4;&#xB780; &#xD65C;&#xC131;&#xD654; polyspermy &#xBC0F; &#xC2DC;&#xC791; &#xB41C; &#xC811;&#xD569; &#xC790; &#xAC1C;&#xBC1C;&#xC744; &#xBC29;&#xC9C0; &#xD558;&#xB294; &#xB370; &#xD544;&#xC694;&#xD55C; &#xC911;&#xC694; &#xD55C; &#xC140;&#xB8F0;&#xB7EC; &#xC774;&#xBCA4;&#xD2B8;&#xC785;&#xB2C8;&#xB2E4;. &#xBAA8;&#xB4E0; &#xB3D9;&#xBB3C; &#xC2DC;&#xD5D8;&#xC5D0;&#xC11C; &#xACC4;&#xB780; &#xD65C;&#xC131;&#xD654;&#xB294; &#xC0C1;&#xC2B9;&#xC758; &#xC218;&#xC815;&#xC5D0;&#xC11C; &#xACC4;&#xB780; cytosol &#xBB34;&#xB8CC; ca(2+)&#xC5D0; &#xC758;&#xD574; elicited. Ca(2+) &#xC0C1;&#xC2B9;&#xC774; &#xB300;&#xB1CC; &#xD53C; &#xC9C8;&#xACFC; &#xB9BD; exocytosis polyspermy&#xB97C; &#xBC29;&#xC9C0; &#xD558;&#xAE30; &#xC704;&#xD574; vitelline &#xBA64;&#xBE0C;&#xB808;&#xC778;&#xC744; &#xC218;&#xC815; &#xD558;&#xB294; &#xB4F1;&#xC758; &#xBAA8;&#xB4E0; &#xD6C4;&#xC18D; &#xACC4;&#xB780; &#xD65C;&#xC131;&#xD654; &#xB2E8;&#xACC4;&#xC5D0; &#xB300; &#xD55C; &#xACB0;&#xC815;&#xC801; &#xC774;&#xB2E4;. Cytosolic Ca(2+) &#xC99D;&#xAC00; &#xC8FC;&#xB85C; inositol 1,4,5 trisphosphate (IP(3))&#xC5D0; &#xC758;&#xD574; &#xC2DC;&#xC791;-Ca(2+) &#xCD9C;&#xC2DC; endoplasmic &#xADF8;&#xBB3C;&#xC5D0;&#xC11C; &#xC911;&#xC7AC;. (3) IP &#xC870;&#xC808;&#xC5D0; &#xAD00;&#xB828; &#xB41C; &#xC720;&#xC804;&#xC790;-&#xACC4;&#xB780; &#xD65C;&#xC131;&#xD654; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xC911;&#xC7AC; Ca(2+) &#xB9B4;&#xB9AC;&#xC2A4; &#xD06C;&#xAC8C; &#xC54C;&#xB824;&#xC9C0;&#xC9C0; &#xC54A;&#xC740; &#xB0A8;&#xC544; &#xC788;&#xB2E4;. &#xC5EC;&#xAE30; &#xC6B0;&#xB9AC;&#xAC00; zebrafish &#xBAA8;&#xC131; &#xD6A8;&#xACFC; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xB300; &#xD55C; &#xBCF4;&#xACE0;&#xC11C; cytosolic Ca(2+) &#xC0C1;&#xC2B9;&#xC5D0; &#xACB0;&#xD568;&#xC774; &#xACE0; &#xD6C4;&#xC18D; &#xBE0C;&#xB86C; &#xBF08; &#xACC4;&#xB780; &#xB300;&#xB1CC; &#xD53C; &#xC9C8;&#xACFC; &#xB9BD; exocytosis &#xC138;&#xD3EC;&#xC9C8; &#xBD84;&#xB9AC;&#xB97C; &#xD3EC;&#xD568; &#xD558;&#xB294; &#xD65C;&#xC131;&#xD654; &#xC774;&#xBCA4;&#xD2B8;. &#xC6B0;&#xB9AC; &#xACC4;&#xB780; &#xD65C;&#xC131;&#xD654; &#xACB0;&#xD568; &#xBE0C;&#xB86C; &#xBF08;&#xC5D0; &#xC81C;&#xACF5; &#xD558;&#xB294; Ca(2+) &#xB610;&#xB294; Ca(2+) &#xB9B4;&#xB9AC;&#xC2A4; &#xBA54;&#xC2E0;&#xC800; IP(3), &#xBE0C;&#xB86C; &#xBF08; IP (3)&#xC758; &#xB808; &#xADE4; &#xB808;&#xC774; &#xD130;&#xB294; &#xC81C;&#xC548;&#xC5D0; &#xC758;&#xD574; &#xAD6C;&#xCD9C; &#xB420; &#xC218; &#xD45C;&#xC2DC;-&#xC911;&#xC7AC; Ca(2+) &#xB9B4;&#xB9AC;&#xC2A4; &#xC218;&#xC815;. &#xD765;&#xBBF8;&#xB86D;&#xAC8C;&#xB3C4;, &#xBE0C;&#xB86C; &#xBF08; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xBC30;&#xC544;&#xB3C4; &#xC9C0; &#xCD95; &#xD615;&#xC131;&#xC744; &#xC704;&#xD55C; &#xC911;&#xC694; &#xD55C; &#xAC83;&#xC73C;&#xB85C; &#xC54C;&#xB824;&#xC9C4; &#xBE44; &#xC870;&#xC9C1;&#xB41C; &#xB300;&#xB1CC; &#xD53C; &#xC9C8;&#xC758; microtubule &#xB124;&#xD2B8;&#xC6CC;&#xD06C;&#xC640; &#xD568;&#xAED8; dorsoventral &#xCD95; &#xD615;&#xC131;&#xC5D0; &#xACB0;&#xD568;&#xC744; &#xD45C;&#xC2DC; &#xD569;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC; &#xC7A5;&#xC560;&#xC778;&#xB41C; microtubule &#xC870;&#xC9C1;&#xC740; &#xC774;&#xC804;&#xC5D0; &#xADF8; &#xD65C;&#xC131;&#xD654; &#xACB0;&#xD568;&#xC5D0;&#xC11C; &#xB300;&#xB1CC; &#xD53C; &#xC9C8;&#xACFC; &#xB9BD; &#xBE44; exocytosed&#xC640; &#xAD00;&#xB828; &#xB41C; &#xC99D;&#xAC70;&#xB97C; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;. &#xBC1D;&#xD600; &#xC704;&#xCE58; &#xBE0C;&#xB86C; &#xBF08; &#xC720;&#xC804;&#xC790;&#xC758; &#xD074;&#xB85C;&#xB2DD; hnRNP &#xC778;&#xCF54;&#xB529; &#xC720;&#xC804;&#xC790;&#xC5D0;&#xC11C; &#xC870; &#xC815;&#xC9C0; codon &#xB098; (&#xB77C;&#xACE0; &#xC5EC;&#xAE30; hnrnp &#xB098;) &#xAE30;&#xCD08;&#xAC00; &#xB418;&#xB294; &#xC774;&#xC0C1;. &#xC6B0;&#xB9AC;&#xC758; &#xC5F0;&#xAD6C;&#xB294; &#xB530;&#xB77C;&#xC11C; hnrnp&#xC758; &#xC911;&#xC694; &#xD55C; &#xC0C8;&#xB85C;&#xC6B4; &#xC5ED;&#xD560; &#xACF5;&#xAC1C; IP (3)&#xC758; &#xAE30;&#xBCF8; &#xACFC;&#xC815;&#xC744; &#xADDC;&#xC81C;&#xC5D0; &#xB098;-Ca(2+) &#xCD9C;&#xC2DC;&#xC5D0; &#xADF8; &#xD65C;&#xC131;&#xD654;&#xC5D0; &#xC911;&#xC7AC;.

hnRNP &#xB0B4;&#xAC00; &#xC6D0;&#xC778; &#xACC4;&#xB780; zebrafish&#xC758; &#xD65C;&#xC131;&#xD654; Ca2 + &#xC2E0;&#xD638;&#xB97C; &#xC0DD;&#xC131; &#xD558;&#xB294; &#xB370; &#xD544;&#xC694;&#xD55C;.

La activaci&#xF3;n del huevo es un evento celular importante necesario para evitar la polispermia e iniciar el desarrollo del cigoto. La activaci&#xF3;n del huevo en todos los animales examinados se desencadena por un aumento de Ca libre (2 +) en el citosol de huevos en la fertilizaci&#xF3;n. Este Ca (2 +) aumento es crucial para todos los pasos subsiguientes de activaci&#xF3;n del huevo, como la exocitosis de los gr&#xE1;nulos corticales, que modifica la membrana vitelina para evitar la polispermia. El Ca citos&#xF3;lico (2 +) aumento est&#xE1; principalmente iniciada por el inositol 1,4,5-trifosfato (IP) (3) mediada por Ca (2 +) de liberaci&#xF3;n desde el ret&#xED;culo endopl&#xE1;smico. Los genes implicados en la regulaci&#xF3;n de la propiedad intelectual (3)-mediada por Ca (2 +) durante la activaci&#xF3;n de la liberaci&#xF3;n de huevos siguen siendo en gran parte desconocido. Aqu&#xED; nos informe sobre un pez cebra mutante materno-efecto, los huesos de Brom, que es defectuoso en el Ca citos&#xF3;lico (2 +) aumento y posteriores eventos de activaci&#xF3;n del huevo, incluyendo exocitosis de los gr&#xE1;nulos corticales y la segregaci&#xF3;n citoplasm&#xE1;tica. Se demuestra que los defectos de activaci&#xF3;n de huevo en los huesos brom puede ser rescatado por proporcionar Ca (2 +) o Ca (2 +) de liberaci&#xF3;n de mensajer&#xED;a IP (3), lo que sugiere que los huesos Brom es un regulador de la propiedad intelectual (3)-mediada por Ca (2 +) en la fertilizaci&#xF3;n de liberaci&#xF3;n. Curiosamente, brom embriones huesos mutantes tambi&#xE9;n muestran defectos en la formaci&#xF3;n de eje dorsoventral acompa&#xF1;ado por una red de microt&#xFA;bulos desorganizado cortical, que se sabe que es crucial para la formaci&#xF3;n eje dorsal. Ofrecemos pruebas de que la organizaci&#xF3;n de los microt&#xFA;bulos deterioro se asocia con organizaciones no exocitado gr&#xE1;nulos corticales del defecto de la activaci&#xF3;n del huevo antes. La clonaci&#xF3;n posicional del gen Brom revela que los huesos de un cod&#xF3;n de parada prematuro en el gen que codifica la que hnRNP (denominado aqu&#xED; como hnRNP I) subyace a las anomal&#xED;as. Nuestros estudios revelan por lo tanto, una nueva e importante funci&#xF3;n de hnRNP I en la regulaci&#xF3;n del proceso fundamental de la propiedad intelectual (3)-mediada por Ca (2 +) en la activaci&#xF3;n de la liberaci&#xF3;n del huevo.

HnRNP que se requiere para generar la se&#xF1;al de Ca2 + que causa la activaci&#xF3;n del huevo en el pez cebra

&#x80B1;&#x808C;&#x8425;&#x517B;&#x4E0D;&#x826F;&#x75C7; (FSHD) &#x662F;&#x5173;&#x4E8E;&#x4EBA;&#x7C7B;&#x67D3;&#x8272;&#x4F53; 4q35 D4Z4 &#x91CD;&#x590D;&#x6536;&#x7F29;&#x4E0E;&#x76F8;&#x5173;&#x8054;&#x3002;&#x8FD9;&#x79CD;&#x9057;&#x4F20;&#x75C5;&#x53D8;&#x4E0D;&#x4F1A;&#x5BFC;&#x81F4;&#x5B8C;&#x5168;&#x4E27;&#x5931;&#x6216;&#x4EFB;&#x4F55;&#x57FA;&#x56E0;&#x7A81;&#x53D8;&#x3002;&#x56E0;&#x6B64;&#xFF0C;FSHD &#x81F4;&#x75C5;&#x673A;&#x5236;&#x662F;&#x5F88;&#x96BE;&#x8FA8;&#x522B;&#x3002;&#x5728;&#x9886;&#x5BFC; FSHD &#x53D1;&#x75C5;&#x673A;&#x7406;&#x6A21;&#x578B;&#xFF0C;D4Z4 &#x6536;&#x7F29;&#x62DF;&#x5BFC;&#x81F4;&#x8868;&#x89C2;&#x9057;&#x4F20;&#x53D8;&#x5316;&#xFF0C;&#x6700;&#x7EC8;&#x589E;&#x52A0;&#x808C;&#x75C5;&#x7684;&#x6F5C;&#x529B;&#x4E0E;&#x57FA;&#x56E0;&#x7684;&#x8868;&#x8FBE;&#x3002;&#x867D;&#x7136;&#x6CA1;&#x6709;&#x57FA;&#x56E0;&#x6700;&#x7EC8;&#x6302;&#x94A9; FSHD &#x53D1;&#x5C55;&#xFF0C;&#x6700;&#x8FD1;&#x7684;&#x8BC1;&#x636E;&#x652F;&#x6301;&#x7F16;&#x7801; D4Z4 DUX4 &#x57FA;&#x56E0;&#x5728; FSHD &#x4E2D;&#x53D1;&#x6325;&#x4F5C;&#x7528;&#x3002;&#x5728;&#x6B64;&#x7814;&#x7A76;&#x4E2D;&#xFF0C;&#x6211;&#x4EEC;&#x7684;&#x76EE;&#x6807;&#x662F;&#x6D4B;&#x8BD5; DUX4 &#x7684;&#x4F53;&#x5185;&#x808C;&#x75C5;&#x6F5C;&#x529B;&#x3002;

DUX4&#xFF0C;&#x5019;&#x9009;&#x57FA;&#x56E0;&#x7684;&#x80B1;&#x808C;&#x8425;&#x517B;&#x4E0D;&#x826F;&#xFF0C;&#x5BFC;&#x81F4;&#x4F53;&#x5185; p53 &#x4F9D;&#x8D56;&#x808C;&#x75C5;&#x3002;

Facioscapulohumeral muscular dystrophy (FSHD) is associated with D4Z4 repeat contraction on human chromosome 4q35. This genetic lesion does not result in complete loss or mutation of any gene. Consequently, the pathogenic mechanisms underlying FSHD have been difficult to discern. In leading FSHD pathogenesis models, D4Z4 contractions are proposed to cause epigenetic changes, which ultimately increase expression of genes with myopathic potential. Although no gene has been conclusively linked to FSHD development, recent evidence supports a role for the D4Z4-encoded DUX4 gene in FSHD. In this study, our objective was to test the in vivo myopathic potential of DUX4.

DUX4, a candidate gene for facioscapulohumeral muscular dystrophy, causes p53-dependent myopathy in vivo.

Facioscapulohumeral muscular dystrophy (FSHD) est li&eacute;e &agrave; la contraction de r&eacute;p&eacute;tition D4Z4 sur le chromosome humain 4q35. Cette l&eacute;sion g&eacute;n&eacute;tique n&#39;entra&icirc;ne pas une perte compl&egrave;te ou la mutation d&#39;un g&egrave;ne. Par cons&eacute;quent, les m&eacute;canismes pathog&egrave;nes FSHD ont &eacute;t&eacute; difficiles &agrave; discerner. Dans les principaux mod&egrave;les de pathogen&egrave;se FSHD, les contractions de D4Z4 sont propos&eacute;es pour provoquer les changements &eacute;pig&eacute;n&eacute;tiques, qui augmentent en fin de compte l&#39;expression de g&egrave;nes ayant un potentiel myopathique. Bien qu&#39;aucun g&egrave;ne n&#39;a &eacute;t&eacute; d&eacute;finitivement li&eacute;e au d&eacute;veloppement de la FSHD, des preuves r&eacute;centes prend en charge un r&ocirc;le pour le g&egrave;ne DUX4 D4Z4 cod&eacute; dans FSHD. Dans cette &eacute;tude, notre objectif &eacute;tait de tester le potentiel myopathique in vivo de DUX4.

DUX4, un g&egrave;ne candidat pour facioscapulohumeral muscular dystrophy, provoque la myopathie p53 d&eacute;pendant in vivo.

Facio-escapulo Muskeldystrophie (FSHD) ist D4Z4 wiederholte Kontraktion auf menschliche Chromosom 4q35 zugeordnet. Diese genetische L&auml;sion f&uuml;hrt nicht zu Verlust oder Mutation des jedes Gen. Folglich wurden die pathogenen Mechanismen FSHD schwer zu erkennen. F&uuml;hrende FSHD Pathogenese Modelle, sind D4Z4 Kontraktionen schlug um epigenetische Ver&auml;nderungen verursachen, die Expression von Genen mit myopathische Potenzial letztlich zu erh&ouml;hen. Obwohl kein Gen schl&uuml;ssig mit FSHD Entwicklung verkn&uuml;pft ist, unterst&uuml;tzt neuere Belege eine Rolle f&uuml;r das Gen DUX4 D4Z4-codierte in FSHD. In dieser Studie war unser Ziel, das in-vivo myopathische Potenzial von DUX4 zu testen.

DUX4, ein Kandidat-gen f&uuml;r facio-escapulo Muskeldystrophie, bewirkt, dass p53-abh&auml;ngige Myopathie in-vivo.

Distrofia muscolare facio scapolo omerale (FSHD) &egrave; associata a contrazione ripetizione D4Z4 sul cromosoma umano 4q35. Questa lesione genetica non comporta perdita completa o mutazione di geni. Di conseguenza, i meccanismi patogeni FSHD sono state difficili da discernere. Nei principali modelli patogenesi FSHD, D4Z4 contrazioni sono proposti a provocare cambiamenti epigenetici, che aumentano in definitiva espressione di geni con potenziale miopatiche. Anche se nessun gene &egrave; stato definitivamente collegato allo sviluppo di FSHD, recenti prove supportano un ruolo per il gene DUX4 con codifica D4Z4 nella FSHD. In questo studio, il nostro obiettivo era di testare il potenziale miopatiche in vivo di DUX4.

Dux4, un gene candidato per la distrofia muscolare facio scapolo omerale, causa miopatia di p53-dependent in vivo.

&#x9854;&#x9762;&#x80A9;&#x7532;&#x4E0A;&#x8155;&#x7B4B;&#x30B8;&#x30B9;&#x30C8;&#x30ED;&#x30D5;&#x30A3;&#x30FC; (FSHD) D4Z4 &#x30EA;&#x30D4;&#x30FC;&#x30C8;&#x53CE;&#x7E2E;&#x306E;&#x3072;&#x3068;&#x67D3;&#x8272;&#x4F53; 4q35 &#x306B;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x907A;&#x4F1D;&#x7684;&#x75C5;&#x5909;&#x3067;&#x306F;&#x3001;&#x5B8C;&#x5168;&#x306A;&#x640D;&#x5931;&#x307E;&#x305F;&#x306F;&#x4EFB;&#x610F;&#x306E;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x7A81;&#x7136;&#x5909;&#x7570;&#x3067;&#x306F;&#x767A;&#x751F;&#x3057;&#x307E;&#x305B;&#x3093;&#x3002;&#x305D;&#x306E;&#x7D50;&#x679C;&#x3001;FSHD &#x306E;&#x75C5;&#x539F;&#x6027;&#x306E;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x3092;&#x898B;&#x5206;&#x3051;&#x308B;&#x306F;&#x56F0;&#x96E3;&#x3067;&#x3057;&#x305F;&#x3002;FSHD &#x75C5;&#x614B;&#x30E2;&#x30C7;&#x30EB;&#x3092;&#x30EA;&#x30FC;&#x30C9;&#x3059;&#x308B;&#x3053;&#x3068;&#x3067;&#x3001;&#x6700;&#x7D42;&#x7684;&#x306B;&#x5897;&#x52A0;&#x7B4B;&#x7D44;&#x7E54;&#x306E;&#x6F5C;&#x5728;&#x7684;&#x907A;&#x4F1D;&#x5B50;&#x767A;&#x73FE;&#x306E;&#x30A8;&#x30D4;&#x30B8;&#x30A7;&#x30CD;&#x30C6;&#x30A3;&#x30C3;&#x30AF;&#x306A;&#x5909;&#x5316;&#x3092;&#x5F15;&#x304D;&#x8D77;&#x3053;&#x3059; D4Z4 &#x306E;&#x53CE;&#x7E2E;&#x3092;&#x63D0;&#x6848;&#x3059;&#x308B;.FSHD &#x958B;&#x767A;&#x306B;&#x907A;&#x4F1D;&#x5B50;&#x6C7A;&#x5B9A;&#x7684;&#x30EA;&#x30F3;&#x30AF;&#x3055;&#x308C;&#x3066;&#x3044;&#x306A;&#x3044;&#x304C;&#x3001;&#x6700;&#x8FD1;&#x306E;&#x8A3C;&#x62E0;&#x306F; D4Z4 &#x30A8;&#x30F3;&#x30B3;&#x30FC;&#x30C9; DUX4 &#x907A;&#x4F1D;&#x5B50; FSHD &#x306E;&#x305F;&#x3081;&#x3001;&#x5F79;&#x5272;&#x3092;&#x30B5;&#x30DD;&#x30FC;&#x30C8;&#x3057;&#x307E;&#x3059;&#x3002;&#x672C;&#x7814;&#x7A76;&#x3067;&#x306F;&#x3001;&#x6211;&#x3005; &#x306E;&#x76EE;&#x7684;&#x306F; DUX4 &#x306E;&#x751F;&#x4F53;&#x5185;&#x7B4B;&#x7D44;&#x7E54;&#x306E;&#x53EF;&#x80FD;&#x6027;&#x3092;&#x30C6;&#x30B9;&#x30C8;&#x3059;&#x308B;&#x306B;&#x306F;&#x3060;&#x3063;&#x305F;&#x3002;

DUX4&#x3001;&#x9854;&#x9762;&#x80A9;&#x7532;&#x4E0A;&#x8155;&#x7B4B;&#x30B8;&#x30B9;&#x30C8;&#x30ED;&#x30D5;&#x30A3;&#x30FC;&#x306E;&#x907A;&#x4F1D;&#x5B50;&#x5019;&#x88DC;&#x751F;&#x4F53;&#x5185;&#x306E; p53 &#x4F9D;&#x5B58;&#x6027;&#x30DF;&#x30AA;&#x30D1;&#x30C1;&#x30FC;&#x304C;&#x767A;&#x751F;&#x3057;&#x307E;&#x3059;&#x3002;

Facioscapulohumeral &#xADFC; &#xC704;&#xCD95; &#xC99D; (FSHD) &#xC778;&#xAC04;&#xC758; &#xC5FC;&#xC0C9;&#xCCB4; 4q35&#xC5D0; D4Z4 &#xBC18;&#xBCF5; &#xC218;&#xCD95;&#xACFC; &#xC5F0;&#xACB0; &#xB429;&#xB2C8;&#xB2E4;. &#xC774; &#xC720;&#xC804;&#xC801; &#xBCD1; &#xBCC0;&#xC758; &#xC644;&#xC804; &#xC190;&#xC2E4; &#xB610;&#xB294; &#xC5B4;&#xB5A4; &#xC720;&#xC804;&#xC790;&#xC758; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xB418;&#xC9C0; &#xC54A;&#xC2B5;&#xB2C8;&#xB2E4;. &#xB530;&#xB77C;&#xC11C;, &#xAE30;&#xBCF8; FSHD &#xBCD1;&#xC6D0; &#xC131; &#xBA54;&#xCEE4;&#xB2C8;&#xC998; &#xBD84;&#xBCC4; &#xD558;&#xAE30; &#xC5B4;&#xB824;&#xC6B4; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;. FSHD pathogenesis &#xBAA8;&#xB378; &#xC120;&#xB3C4;&#xC5D0;&#xC11C; D4Z4 &#xC218;&#xCD95; &#xAD81;&#xADF9;&#xC801;&#xC73C;&#xB85C; &#xC99D;&#xAC00; &#xC2DD; myopathic &#xC7A0;&#xC7AC;&#xB825;&#xC744; &#xAC00;&#xC9C4; &#xC720;&#xC804;&#xC790;&#xC758; epigenetic &#xBCC0;&#xD654;&#xB97C; &#xC77C;&#xC73C;&#xD0A4;&#xB294; &#xAC83;&#xC73C;&#xB85C; &#xC81C;&#xC548; &#xD569;&#xB2C8;&#xB2E4;. &#xC544;&#xBB34; &#xC720;&#xC804;&#xC790; FSHD &#xAC1C;&#xBC1C;&#xC5D0; &#xACB0;&#xC815;&#xC801;&#xC73C;&#xB85C; &#xC5F0;&#xACB0; &#xB418;&#xC5B4; &#xC788;&#xC9C0;&#xB9CC; &#xCD5C;&#xADFC;&#xC758; &#xC99D;&#xAC70; FSHD&#xC5D0; d4z4&#xB85C; &#xC778;&#xCF54;&#xB529;&#xB41C; DUX4 &#xC720;&#xC804;&#xC790;&#xC5D0; &#xB300; &#xD55C; &#xC5ED;&#xD560;&#xC744; &#xC9C0;&#xC6D0; &#xD569;&#xB2C8;&#xB2E4;. &#xC774; &#xC5F0;&#xAD6C;&#xC5D0;&#xC11C; &#xC6B0;&#xB9AC;&#xC758; &#xBAA9;&#xD45C;&#xB294; dux4&#xC758; in vivo myopathic &#xC7A0;&#xC7AC;&#xB825;&#xC744; &#xD14C;&#xC2A4;&#xD2B8; &#xD588;&#xB2E4;.

DUX4, facioscapulohumeral &#xADFC; &#xC704;&#xCD95; &#xC99D;&#xC5D0; &#xB300; &#xD55C; &#xD6C4;&#xBCF4; &#xC720;&#xC804;&#xC790; p53 &#xC885;&#xC18D; myopathy vivo&#xC5D0;&#xC11C; &#xBC1C;&#xC0DD;&#xD569;&#xB2C8;&#xB2E4;.

La distrofia muscular facioescapulohumeral (FSHD) se asocian con la contracci&oacute;n repetici&oacute;n D4Z4 en el cromosoma humano 4q35. Esta lesi&oacute;n gen&eacute;tica no lleva a la p&eacute;rdida completa o mutaci&oacute;n de cualquier gen. En consecuencia, los mecanismos patog&eacute;nicos subyacentes FSHD han sido dif&iacute;ciles de discernir. En los principales modelos de la patogenesia FSHD, contracciones D4Z4 se proponen para causar cambios epigen&eacute;ticos, que en &uacute;ltima instancia aumentan la expresi&oacute;n de genes con potencial miop&aacute;ticos. Aunque ning&uacute;n gen ha sido concluyente relacionado con desarrollo de FSHD, evidencia reciente apoya el papel del gen DUX4 D4Z4 codificados en FSHD. En este estudio, nuestro objetivo era probar el potencial miop&aacute;ticos in vivo de DUX4.

DUX4, un gen candidato para la distrofia muscular facioescapulohumeral, causa miopat&iacute;a dependiente de p53 en vivo.

La v&iacute;a de Hedgehog (Hh) es evolutivamente conservada y juega un papel fundamental durante el desarrollo embrionario y la homeostasis de tejido adulto. Se&ntilde;alizaci&oacute;n defectuosa de Hh se ha relacionado con una amplia gama de c&aacute;nceres y malformaciones cong&eacute;nitas. HH familias prote&iacute;nas regulan la expresi&oacute;n de sus genes diana mediante el control de procesamiento proteol&iacute;tico y la funci&oacute;n de activaci&oacute;n transcripcional de factores de transcripci&oacute;n de Gli. Aunque el Reglamento de Hh-dependiente de Gli ha sido estudiado ampliamente, otros mecanismos normativos de Gli siguen siendo relativamente poco apreciados. Presentamos nuestra identificaci&oacute;n de una novela de se&ntilde;alizaci&oacute;n cascada que controla la estabilidad de las prote&iacute;nas de Gli. Esta cascada est&aacute; constituido por Daz interacci&oacute;n prote&iacute;na 1 (Dzip1), case&iacute;na quinasa 2 (CK2), y B56 que contiene prote&iacute;na fosfatasa 2As (PP2As). Proporcionamos evidencia que Dzip1 est&aacute; involucrado en un nuevo camino de facturaci&oacute;n Gli. Mostramos que CK2 fosforila directamente Dzip1 en cuatro residuos de serina, Ser-664/665/706/714. PP2As que contienen B56, mediante el enlace a un dominio situado entre residuo de amino&aacute;cido 474 y 550 de Dzip1, dephosphorylate Dzip1 en estos sitios CK2. Nuestro an&aacute;lisis de mutag&eacute;nesis m&aacute;s demuestran que la forma unphosphorylatable de Dzip1 es m&aacute;s potente en la promoci&oacute;n de facturaci&oacute;n de Gli. Constantemente, encontramos que la estabilidad de las prote&iacute;nas Gli fue disminuida a la inhibici&oacute;n de la CK2 y aumentada por la inhibici&oacute;n de PP2As que contienen B56. As&iacute;, la fosforilaci&oacute;n reversible de Dzip1, que es controlada por la acci&oacute;n antagonista de la CK2 y PP2As que contienen B56, tiene un impacto importante en la estabilidad de los factores de transcripci&oacute;n de Gli y se&ntilde;alizaci&oacute;n Hh.

La acci&oacute;n antag&oacute;nica de B56 que contienen prote&iacute;na fosfatasa 2As y case&iacute;na quinasa 2 controla la fosforilaci&oacute;n y la funci&oacute;n de facturaci&oacute;n de Gli de Daz interacci&oacute;n prote&iacute;na 1.

&#x523A;&#x732C; (Hh) &#x901A;&#x8DEF;&#x8FDB;&#x5316;&#x5B88;&#x6052;&#x548C;&#x80DA;&#x80CE;&#x53D1;&#x80B2;&#x548C;&#x6210;&#x4EBA;&#x7EC4;&#x7EC7;&#x7A33;&#x6001;&#x671F;&#x95F4;&#x626E;&#x6F14;&#x5173;&#x952E;&#x89D2;&#x8272;&#x3002;&#x6709;&#x7F3A;&#x9677; Hh &#x4FE1;&#x53F7;&#x5DF2;&#x94FE;&#x63A5;&#x5230;&#x5E7F;&#x6CDB;&#x7684;&#x51FA;&#x751F;&#x7F3A;&#x9677;&#x548C;&#x764C;&#x75C7;&#x3002;Hh &#x5BB6;&#x65CF;&#x86CB;&#x767D;&#x8C03;&#x8282;&#x86CB;&#x767D;&#x6C34;&#x89E3;&#x5904;&#x7406;&#x63A7;&#x5236;&#x548C; Gli &#x8F6C;&#x5F55;&#x56E0;&#x5B50;&#x7684;&#x8F6C;&#x5F55;&#x6FC0;&#x6D3B;&#x529F;&#x80FD;&#x901A;&#x8FC7;&#x5176;&#x4E0B;&#x6E38;&#x9776;&#x57FA;&#x56E0;&#x7684;&#x8868;&#x8FBE;&#x3002;&#x867D;&#x7136; Gli Hh &#x4F9D;&#x8D56;&#x8C03;&#x8282;&#x4E86;&#x5E7F;&#x6CDB;&#x7684;&#x7814;&#x7A76;&#xFF0C;&#x5176;&#x4ED6; Gli &#x76D1;&#x7BA1;&#x673A;&#x5236;&#x4ECD;&#x7136;&#x76F8;&#x5BF9;&#x8D4F;&#x8BC6;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x62A5;&#x544A;&#x6211;&#x4EEC;&#x9274;&#x5B9A;&#x7684;&#x4E00;&#x79CD;&#x65B0;&#x578B;&#x4FE1;&#x53F7;&#x63A7;&#x5236;&#x7684; Gli &#x86CB;&#x767D;&#x7A33;&#x5B9A;&#x6027;&#x7684;&#x7EA7;&#x8054;&#x3002;&#x6B64;&#x7EA7;&#x8054;&#x5305;&#x542B; Daz &#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x86CB;&#x767D; 1 (Dzip1)&#x3001; &#x916A;&#x86CB;&#x767D;&#x6FC0;&#x9176; 2 &#xFF08;CK2&#xFF09; &#x548C; B56 &#x542B;&#x86CB;&#x767D;&#x78F7;&#x9178;&#x9176; 2As (PP2As)&#x3002;&#x6211;&#x4EEC;&#x63D0;&#x4F9B;&#x8BC1;&#x636E; Dzip1 &#x53C2;&#x4E0E;&#x65B0;&#x578B; Gli &#x8425;&#x4E1A;&#x989D;&#x901A;&#x8DEF;&#x3002;&#x6211;&#x4EEC;&#x5C55;&#x793A; CK2 &#x76F4;&#x63A5;&#x5728;&#x56DB;&#x4E2A;&#x4E1D;&#x6C28;&#x9178;&#x6B8B;&#x7559;&#xFF0C;Ser-664/665/706/714 phosphorylates Dzip1&#x3002;B56 &#x5305;&#x542B; PP2As&#xFF0C;&#x7ED1;&#x5B9A;&#x5230;&#x4E00;&#x4E2A;&#x57DF;&#xFF0C;&#x4F4D;&#x4E8E;&#x6C28;&#x57FA;&#x9178;&#x6B8B;&#x57FA; 474 &#x548C; Dzip1&#xFF0C;550 &#x4E4B;&#x95F4;&#x901A;&#x8FC7;&#x5176;&#x5BF9;&#x8FD9;&#x4E9B; CK2 &#x7F51;&#x7AD9; Dzip1&#x3002;&#x6211;&#x4EEC;&#x7684;&#x7A81;&#x53D8;&#x5206;&#x6790;&#x8FDB;&#x4E00;&#x6B65;&#x6F14;&#x793A; unphosphorylatable Dzip1 &#x7684;&#x5F62;&#x5F0F;&#x662F;&#x5728;&#x4FC3;&#x8FDB; Gli &#x8425;&#x4E1A;&#x989D;&#x66F4;&#x6709;&#x529B;&#x3002;&#x59CB;&#x7EC8;&#x5982;&#x4E00;&#xFF0C;&#x6211;&#x4EEC;&#x53D1;&#x73B0; Gli &#x86CB;&#x767D;&#x7684;&#x7A33;&#x5B9A;&#x6027;&#x662F; CK2 &#x6291;&#x5236;&#x540E;&#x4E0B;&#x964D;&#x548C;&#x4E0A;&#x5347; B56 &#x5305;&#x542B; PP2As &#x7684;&#x6291;&#x5236;&#x4F5C;&#x7528;&#x3002;&#x56E0;&#x6B64;&#xFF0C;&#x53EF;&#x9006;&#x78F7;&#x9178;&#x5316;&#x7684; Dzip1&#xFF0C;&#x5B83;&#x7531; CK2 &#x548C; B56 &#x5305;&#x542B; PP2As &#x7684;&#x62EE;&#x6297;&#x4F5C;&#x7528;&#x63A7;&#x5236;&#xFF0C;&#x5BF9; Gli &#x8F6C;&#x5F55;&#x56E0;&#x5B50;&#x548C; Hh &#x4FE1;&#x53F7;&#x7684;&#x7A33;&#x5B9A;&#x6709;&#x91CD;&#x8981;&#x7684;&#x5F71;&#x54CD;&#x3002;

B56 &#x542B;&#x86CB;&#x767D;&#x78F7;&#x9178;&#x9176; 2As &#x548C;&#x916A;&#x86CB;&#x767D;&#x6FC0;&#x9176; 2 &#x7684;&#x62EE;&#x6297;&#x4F5C;&#x7528;&#x63A7;&#x5236;&#x7684;&#x78F7;&#x9178;&#x5316;&#x4E0E; Daz &#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x86CB;&#x767D; 1 Gli &#x8425;&#x4E1A;&#x989D;&#x529F;&#x80FD;&#x3002;

The Hedgehog (Hh) pathway is evolutionarily conserved and plays critical roles during embryonic development and adult tissue homeostasis. Defective Hh signaling has been linked to a wide range of birth defects and cancers. Hh family proteins regulate the expression of their downstream target genes through the control of proteolytic processing and the transcriptional activation function of Gli transcription factors. Although Hh-dependent regulation of Gli has been studied extensively, other Gli regulatory mechanisms remain relatively unappreciated. Here we report our identification of a novel signaling cascade that controls the stability of Gli proteins. This cascade consists of Daz interacting protein 1 (Dzip1), casein kinase 2 (CK2), and B56 containing protein phosphatase 2As (PP2As). We provide evidence that Dzip1 is involved in a novel Gli turnover pathway. We show that CK2 directly phosphorylates Dzip1 at four serine residues, Ser-664/665/706/714. B56-containing PP2As, through binding to a domain located between amino acid residue 474 and 550 of Dzip1, dephosphorylate Dzip1 on these CK2 sites. Our mutagenesis analysis further demonstrates that the unphosphorylatable form of Dzip1 is more potent in promoting Gli turnover. Consistently, we found that the stability of Gli proteins was decreased upon CK2 inhibition and increased by inhibition of B56-containing PP2As. Thus, reversible phosphorylation of Dzip1, which is controlled by the antagonistic action of CK2 and B56-containing PP2As, has an important impact on the stability of Gli transcription factors and Hh signaling.

The antagonistic action of B56-containing protein phosphatase 2As and casein kinase 2 controls the phosphorylation and Gli turnover function of Daz interacting protein 1.

La voie Hedgehog (Hh) est conserv&eacute;e &eacute;volutives et joue un r&ocirc;le crucial au cours du d&eacute;veloppement embryonnaire et l&#39;hom&eacute;ostasie tissulaire adulte. Signalisation de Hh d&eacute;fectueux a &eacute;t&eacute; li&eacute;e &agrave; un large &eacute;ventail de cancers et de malformations cong&eacute;nitales. Prot&eacute;ines de la famille HH r&eacute;gulent l&#39;expression de leurs g&egrave;nes cibles en aval &agrave; travers le contr&ocirc;le de la prot&eacute;olyse et la fonction d&#39;activation de la transcription des facteurs de transcription Gli. Bien que le r&egrave;glement Hh d&eacute;pendant de Gli a &eacute;t&eacute; largement &eacute;tudi&eacute;e, autres m&eacute;canismes r&eacute;gulateurs Gli restent relativement m&eacute;connus. Nous pr&eacute;sentons notre identification of a novel signaling cascade qui contr&ocirc;le la stabilit&eacute; des prot&eacute;ines Gli. Cette cascade se compose de prot&eacute;ines qui interagissent Daz 1 (Dzip1), cas&eacute;ine kinase 2 (CK2), et B56 contenant la prot&eacute;ine phosphatase 2As (PP2As). Nous d&eacute;montrons que Dzip1 est impliqu&eacute; dans une nouvelle voie de chiffre d&#39;affaires de Gli. Nous montrons que CK2 phosphoryle directement Dzip1 &agrave; quatre r&eacute;sidus de s&eacute;rine, Ser-664/665/706/714. B56 contenant PP2As, via la liaison &agrave; un domaine situ&eacute; entre les r&eacute;sidus d&#39;acide amin&eacute; 474 et 550 de Dzip1, d&eacute;phosphorylent Dzip1 sur ces sites de CK2. Notre analyse de mutagen&egrave;se plus d&eacute;montre que la forme Pak1 de Dzip1 est plus puissante dans la promotion du chiffre d&#39;affaires de Gli. R&eacute;guli&egrave;rement, nous avons constat&eacute; que la stabilit&eacute; des prot&eacute;ines Gli a &eacute;t&eacute; diminu&eacute;e sur l&#39;inhibition de la CK2 et augment&eacute;e par inhibition de la PP2As contenant B56. Ainsi, la phosphorylation r&eacute;versible des Dzip1, qui est contr&ocirc;l&eacute;e par l&#39;action antagoniste de CK2 et B56 contenant PP2As, a un impact important sur la stabilit&eacute; des facteurs de transcription Gli et signalisation de Hh.

L&#39;action antagoniste du B56 contenant prot&eacute;ine phosphatase 2As et cas&eacute;ine kinase 2 contr&ocirc;le la phosphorylation et la fonction de chiffre d&#39;affaires de Gli de Daz interaction prot&eacute;ine 1.

Der Igel (Hh)-Stoffwechselweg ist evolution&auml;r konserviert und kritische Rollen w&auml;hrend der embryonalen Entwicklung und Erwachsenen Gewebe Hom&ouml;ostase spielt. Defekte Hh-Signalisierung wird auf ein breites Spektrum von Missbildungen und Krebserkrankungen assoziiert. HH Familie Proteine regulieren den Ausdruck ihrer nachgeschalteten Zielgene durch die Kontrolle der proteolytische Verarbeitung und die transkriptionelle Aktivierungsfunktion des Gli Transkriptionsfaktoren. Obwohl Hh-abh&auml;ngige Regelung der Gli ausgiebig studiert hat, bleiben andere Gli Regulierungsmechanismen relativ unbeachtet. Hier berichten wir &uuml;ber unsere Kennzeichnung eines Romans Signalisierung Kaskade, die die Stabilit&auml;t des Gli Proteinen steuert. Dieser Kaskade besteht aus Daz zusammenwirkendes Protein 1 (Dzip1), Kasein-Kinase 2 (CK2), B56, enthalten Protein Phosphatase 2As (PP2As). Wir belegen, dass eine neuartige Gli Umsatz Weg Dzip1 beteiligt ist. Wir zeigen, dass CK2 direkt Dzip1 an vier Serin R&uuml;ckst&auml;nde, Ser-664/665/706/714 phosphorylates. B56-haltigen PP2As, dephosphorylate durch Bindung an eine Dom&auml;ne befindet sich zwischen Aminos&auml;ure R&uuml;ckstand 474 und 550 von Dzip1, Dzip1 auf diesen CK2-Seiten. Weiter unsere Mutagenese-Analyse zeigt, dass die unphosphorylatable Form der Dzip1 st&auml;rker f&ouml;rdern Gli Umsatz ist. Konsequent, fanden wir, dass die Stabilit&auml;t der Gli Proteine wurde auf CK2 Hemmung verringert und durch Hemmung der B56-haltigen PP2As erh&ouml;ht. Reversible Phosphorylierung von Dzip1, der durch die Antagonistische Wirkung der CK2 und B56-haltigen PP2As kontrolliert wird, hat somit, erhebliche Auswirkungen auf die Stabilit&auml;t des Gli Transkriptionsfaktoren und Hh signalisieren.

Die Antagonistische Wirkung der B56-haltige Protein Phosphatase 2As und Kasein Kinase 2 steuert die Phosphorylierung und Gli Umsatz Funktion Daz interagierenden Proteins 1.

Il pathway Hedgehog (Hh) &egrave; evolutivamente conservato e svolge ruoli importanti durante lo sviluppo embrionale e l&#39;omeostasi dei tessuti adulti. Segnalazione Hh difettoso &egrave; stato collegato a un&#39;ampia gamma di tumori e malformazioni alla nascita. HH famiglie proteine regolano l&#39;espressione dei loro geni bersaglio a valle attraverso il controllo del trattamento proteolitico e la funzione di attivazione trascrizionale dei fattori di trascrizione Gli. Sebbene il regolamento Hh-dipendente di Gli &egrave; stato studiato approfonditamente, altri meccanismi regolatori Gli rimangono relativamente misconosciuti. Riportiamo la nostra identificazione di un romanzo di cascata che controlla la stabilit&agrave; delle proteine Gli di segnalazione. Questa cascata &egrave; costituito da Daz proteina d&#39;interazione 1 (Dzip1), chinasi 2 (CK2), della caseina e B56 contenenti proteine fosfatasi 2As (PP2As). Forniamo prove che Dzip1 &egrave; coinvolto in un percorso di fatturato Gli romanzo. Mostriamo che CK2 fosforila direttamente Dzip1 a quattro residui di Serina, Ser-664/665/706/714. B56-contenenti PP2As, tramite l&#39;associazione a un dominio che si trova tra i residui dell&#39;amminoacido 474 e 550 di Dzip1, dephosphorylate Dzip1 su questi siti CK2. La nostra analisi di mutagenesi ulteriormente dimostra che la forma unphosphorylatable di Dzip1 &egrave; pi&ugrave; potente nel promuovere Gli fatturato. Coerentemente, abbiamo trovato che la stabilit&agrave; delle proteine Gli era diminuita su CK2 inibizione e aumentata tramite inibizione di PP2As contenenti B56. Cos&igrave;, la fosforilazione reversibile di Dzip1, che &egrave; controllata dall&#39;azione antagonista di CK2 e B56-contenenti PP2As, ha un impatto importante sulla stabilit&agrave; dei fattori di trascrizione Gli e segnalazione Hh.

L&#39;azione antagonista di B56-contenenti proteine fosfatasi 2As e caseina chinasi 2 controlla la fosforilazione e la funzione fatturato Gli di Daz proteina d&#39;interazione 1.

&#x30D8;&#x30C3;&#x30B8;&#x30DB;&#x30C3;&#x30B0; (Hh) &#x7D4C;&#x8DEF;&#x306F;&#x9032;&#x5316;&#x7684;&#x306B;&#x4FDD;&#x5B58;&#x3055;&#x308C;&#x3066;&#x3001;&#x80DA;&#x304A;&#x3088;&#x3073;&#x6210;&#x4F53;&#x7D44;&#x7E54;&#x6052;&#x5E38;&#x6027;&#x4E2D;&#x306B;&#x91CD;&#x8981;&#x306A;&#x5F79;&#x5272;&#x3092;&#x679C;&#x305F;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x6B20;&#x9665; Hh &#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x306F;&#x3001;&#x5148;&#x5929;&#x6027;&#x6B20;&#x640D;&#x75C7;&#x3084;&#x764C;&#x306E;&#x5E83;&#x3044;&#x7BC4;&#x56F2;&#x306B;&#x30EA;&#x30F3;&#x30AF;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;Hh &#x30D5;&#x30A1;&#x30DF;&#x30EA;&#x30FC;&#x86CB;&#x767D;&#x8CEA;&#x86CB;&#x767D;&#x8CEA;&#x5206;&#x89E3;&#x51E6;&#x7406;&#x306E;&#x5236;&#x5FA1;&#x3068; Gli &#x8EE2;&#x5199;&#x56E0;&#x5B50;&#x306E;&#x8EE2;&#x5199;&#x6D3B;&#x6027;&#x5316;&#x95A2;&#x6570;&#x3092;&#x901A;&#x3058;&#x3066;&#x4E0B;&#x6D41;&#x30BF;&#x30FC;&#x30B2;&#x30C3;&#x30C8;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x767A;&#x73FE;&#x3092;&#x8ABF;&#x7BC0;&#x3057;&#x307E;&#x3059;&#x3002;Gli Hh &#x4F9D;&#x5B58;&#x7684;&#x306A;&#x8ABF;&#x7BC0;&#x304C;&#x5E83;&#x304F;&#x7814;&#x7A76;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x304C;&#x3001;&#x4ED6; Gli &#x8ABF;&#x7BC0;&#x6A5F;&#x69CB;&#x306F;&#x6BD4;&#x8F03;&#x7684;&#x5275;&#x308B;&#x6B8B;&#x308B;&#x3002;&#x79C1;&#x305F;&#x3061;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054; Gli &#x306E;&#x86CB;&#x767D;&#x8CEA;&#x306E;&#x5B89;&#x5B9A;&#x6027;&#x3092;&#x5236;&#x5FA1;&#x3059;&#x308B;&#x30AB;&#x30B9;&#x30B1;&#x30FC;&#x30C9;&#x5C0F;&#x8AAC;&#x306E;&#x540C;&#x5B9A;&#x3092;&#x5831;&#x544A;&#x3057;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x30AB;&#x30B9;&#x30B1;&#x30FC;&#x30C9; Daz &#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x306E;&#x86CB;&#x767D;&#x8CEA;&#x306E; 1 &#xFF08;Dzip1&#xFF09; &#x5E74;&#x30AB;&#x30BC;&#x30A4;&#x30F3;&#x30AD;&#x30CA;&#x30FC;&#x30BC; 2 (CK2&#xFF09; &#x304B;&#x3089;&#x6210;&#x308A;&#x307E;&#x3059;&#x3001;B56 &#x3092;&#x542B;&#x3080;&#x86CB;&#x767D;&#x8CEA;&#x30DB;&#x30B9;&#x30D5;&#x30A1;&#x30BF;&#x30FC;&#x30BC; 2 as (PP2As)&#x3002;Dzip1 &#x65B0;&#x898F; Gli &#x96E2;&#x8077;&#x7D4C;&#x8DEF;&#x306B;&#x95A2;&#x4E0E;&#x3057;&#x3066;&#x3044;&#x308B;&#x3053;&#x3068;&#x306E;&#x8A3C;&#x62E0;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x307E;&#x3059;&#x3002;CK2 &#x304C; 4 &#x30BB;&#x30EA;&#x30F3;&#x6B8B;&#x57FA;&#x3001;Ser-664/665/706/714 Dzip1 &#x3092;&#x76F4;&#x63A5;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;B56 &#x3092;&#x542B;&#x3080; PP2As &#x30A2;&#x30DF;&#x30CE;&#x9178;&#x6B8B; 474 &#x3068; Dzip1&#x3001;550 &#x306E;&#x9593;&#x306B;&#x5B58;&#x5728;&#x3059;&#x308B;&#x30C9;&#x30E1;&#x30A4;&#x30F3;&#x3078;&#x306E;&#x30D0;&#x30A4;&#x30F3;&#x30C7;&#x30A3;&#x30F3;&#x30B0;&#x3092;&#x4ECB;&#x3057;&#x3066;&#x3053;&#x308C;&#x3089; CK2 &#x306E;&#x30B5;&#x30A4;&#x30C8;&#x4E0A;&#x306E; Dzip1 &#x3092; dephosphorylate &#x3057;&#x307E;&#x3059;&#x3002;&#x79C1;&#x305F;&#x3061;&#x306E;&#x5909;&#x7570;&#x89E3;&#x6790;&#x3092;&#x3055;&#x3089;&#x306B; Dzip1 &#x306E; unphosphorylatable &#x30D5;&#x30A9;&#x30FC;&#x30E0;&#x304C; Gli &#x56DE;&#x8EE2;&#x3092;&#x4FC3;&#x9032;&#x3059;&#x308B;&#x3053;&#x3068;&#x3067;&#x3088;&#x308A;&#x5F37;&#x529B;&#x306A;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x3066;&#x304F;&#x3060;&#x3055;&#x3044;&#x3002;&#x4E00;&#x8CAB;&#x3057;&#x3066;&#x3001;Gli &#x306E;&#x86CB;&#x767D;&#x8CEA;&#x306E;&#x5B89;&#x5B9A;&#x6027;&#x304C; CK2 &#x963B;&#x5BB3;&#x8981;&#x6E1B;&#x5C11;&#x3055;&#x308C; B56 &#x3092;&#x542B;&#x3080; PP2As &#x306E;&#x963B;&#x5BB3;&#x306B;&#x3088;&#x3063;&#x3066;&#x5897;&#x52A0;&#x3092;&#x767A;&#x898B;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x3001;CK2 &#x3068; B56 &#x3092;&#x542B;&#x3080; PP2As &#x306E;&#x62EE;&#x6297;&#x4F5C;&#x7528;&#x306B;&#x3088;&#x3063;&#x3066;&#x5236;&#x5FA1;&#x3055;&#x308C;&#x307E;&#x3059;&#x3001;Dzip1 &#x306E;&#x30EA;&#x30D0;&#x30FC;&#x30B7;&#x30D6;&#x30EB;&#x306E;&#x30EA;&#x30F3;&#x9178;&#x5316;&#x306F;&#x3001;Gli &#x8EE2;&#x5199;&#x56E0;&#x5B50;&#x3068; Hh &#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x306E;&#x5B89;&#x5B9A;&#x6027;&#x306B;&#x95A2;&#x3059;&#x308B;&#x91CD;&#x8981;&#x306A;&#x5F71;&#x97FF;&#x3092;&#x4E0E;&#x3048;&#x307E;&#x3059;&#x3002;

B56 &#x3092;&#x542B;&#x3080;&#x86CB;&#x767D;&#x8CEA;&#x30DB;&#x30B9;&#x30D5;&#x30A1;&#x30BF;&#x30FC;&#x30BC; 2 as &#x3068;&#x30AB;&#x30BC;&#x30A4;&#x30F3; &#x30AD;&#x30CA;&#x30FC;&#x30BC; 2 &#x306E;&#x62EE;&#x6297;&#x4F5C;&#x7528;&#x306E;&#x30EA;&#x30F3;&#x9178;&#x5316;&#x3068; Daz &#x306E;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x306E;&#x86CB;&#x767D;&#x8CEA; 1 Gli &#x58F2;&#x308A;&#x4E0A;&#x3052;&#x9AD8;&#x6A5F;&#x80FD;&#x3092;&#x5236;&#x5FA1;&#x3057;&#x307E;&#x3059;&#x3002;

&#xACE0;&#xC2B4;&#xB3C4;&#xCE58; (Hh) &#xD1B5;&#xB85C; &#xC9C4;&#xD654; &#xC801;&#xC73C;&#xB85C; &#xBCF4;&#xC874; &#xD558; &#xACE0; &#xBC30;&#xC544; &#xAC1C;&#xBC1C; &#xBC0F; &#xC131;&#xC778; &#xC870;&#xC9C1;&#xC758; &#xD56D;&#xC0C1;&#xC131; &#xC911; &#xC911;&#xC694; &#xD55C; &#xC5ED;&#xD560;&#xC744; &#xD55C;&#xB2E4;. &#xACB0;&#xD568;&#xC774; &#xC788;&#xB294; Hh &#xC2E0;&#xD638; &#xB2E4;&#xC591; &#xD55C; &#xAE30;&#xD615;&#xACFC; &#xC554;&#xC5D0; &#xC5F0;&#xACB0; &#xB418;&#xC5B4;&#xC788;&#xB2E4;. Hh &#xAC00;&#xC871; &#xB2E8;&#xBC31;&#xC9C8; &#xC870;&#xC808; &#xB2E8;&#xBC31;&#xC9C8; &#xCC98;&#xB9AC; &#xC81C;&#xC5B4; &#xBC0F; Gli &#xB179;&#xC74C; &#xBC29;&#xC1A1; &#xC694;&#xC778;&#xC758; transcriptional &#xD65C;&#xC131;&#xD654; &#xAE30;&#xB2A5;&#xC744; &#xD1B5;&#xD574; &#xB2E4;&#xC6B4;&#xC2A4;&#xD2B8;&#xB9BC; &#xB300;&#xC0C1; &#xC720;&#xC804;&#xC790;&#xC758; &#xD45C;&#xD604;. Gli&#xC758; Hh &#xC885;&#xC18D; &#xADDC;&#xC81C; &#xAD11;&#xBC94;&#xC704; &#xD558; &#xAC8C; &#xACF5;&#xBD80; &#xD558;&#xACE0;&#xC788;&#xB2E4;, &#xD558;&#xC9C0;&#xB9CC; &#xB2E4;&#xB978; Gli &#xADDC;&#xC81C; &#xBA54;&#xCEE4;&#xB2C8;&#xC998; &#xC0C1;&#xB300;&#xC801;&#xC73C;&#xB85C; &#xC9C4;&#xAC00; &#xB0A8;&#xC544; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC5EC;&#xAE30;&#xC11C; &#xC6B0;&#xB9AC;&#xAC00; &#xC6B0;&#xB9AC;&#xC758; &#xC18C;&#xC124; &#xCE90;&#xC2A4;&#xCF00;&#xC774;&#xB4DC; Gli &#xB2E8;&#xBC31;&#xC9C8;&#xC758; &#xC548;&#xC815;&#xC131;&#xC744; &#xC81C;&#xC5B4; &#xD558;&#xB294; &#xC2E0;&#xD638; &#xC2DD;&#xBCC4; &#xBCF4;&#xACE0;&#xC11C;. &#xC774; &#xBAA8;&#xB450; &#xC774;&#xB8E8;&#xC5B4;&#xC838; Daz &#xC0C1;&#xD638; &#xC791;&#xC6A9; &#xB2E8;&#xBC31;&#xC9C8; 1 (Dzip1), &#xCE74; &#xC138; &#xC778; &#xB2C8; 2 (CK2) B56 &#xB2E8;&#xBC31;&#xC9C8; &#xC778;&#xC0B0; &#xAC00;&#xC218;&#xBD84;&#xD574; &#xD6A8;&#xC18C; 2as&#xB97C; &#xD3EC;&#xD568; &#xD558; &#xACE0; (PP2As). &#xC6B0;&#xB9AC; Dzip1 &#xC18C;&#xC124; Gli &#xB9E4;&#xCD9C; &#xD1B5;&#xB85C;&#xC5D0; &#xAD00;&#xC5EC; &#xD558;&#xB294; &#xC99D;&#xAC70;&#xB97C; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC; CK2 &#xC9C1;&#xC811; &#xB124; &#xB5A0;&#xB4E4;&#xACE0; &#xC794;&#xB958;&#xBB3C;, Ser-664/665/706/714 &#xBD84;&#xC5D0;&#xC11C; Dzip1 phosphorylates &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;. B56 &#xD3EC;&#xD568; &#xB41C; PP2As &#xC544;&#xBBF8;&#xB178;&#xC0B0; &#xC794;&#xB958;&#xBB3C; 474 &#xBC0F; Dzip1, 550 &#xC0AC;&#xC774; &#xC704;&#xCE58;&#xD55C; &#xB3C4;&#xBA54;&#xC778;&#xC5D0; &#xBC14;&#xC778;&#xB529;&#xC744; &#xD1B5;&#xD574; &#xC774;&#xB7EC;&#xD55C; CK2 &#xC0AC;&#xC774;&#xD2B8;&#xC5D0; Dzip1 dephosphorylate. Mutagenesis &#xBD84;&#xC11D; &#xCD94;&#xAC00; unphosphorylatable &#xD615;&#xD0DC;&#xC758; Dzip1 Gli &#xD68C;&#xC804;&#xC728;&#xC744; &#xD64D;&#xBCF4;&#xC5D0; &#xB354; &#xAC15;&#xB825;&#xD55C; &#xAC83;&#xC744; &#xBCF4;&#xC5EC; &#xC90D;&#xB2C8;&#xB2E4;. &#xC9C0;&#xC18D;&#xC801;&#xC73C;&#xB85C;, &#xC6B0;&#xB9AC;&#xB294; Gli &#xB2E8;&#xBC31;&#xC9C8;&#xC758; &#xC548;&#xC815;&#xC131; CK2 &#xC5B5;&#xC81C;&#xC5D0; &#xB530;&#xB77C; &#xAC10;&#xC18C; &#xD558; &#xACE0; B56 &#xD3EC;&#xD568; &#xB41C; pp2as&#xC758; &#xC800;&#xD574;&#xC5D0; &#xC758;&#xD574; &#xC99D;&#xAC00; &#xD588;&#xB2E4; &#xBC1C;&#xACAC;. &#xB530;&#xB77C;&#xC11C;, CK2 B56 &#xD3EC;&#xD568; &#xB41C; pp2as&#xC758; &#xB300;&#xB9BD; &#xD589;&#xB3D9;&#xC5D0; &#xC758;&#xD574; &#xC81C;&#xC5B4; &#xB418;&#xB294; dzip1&#xC758; &#xAC00;&#xC5ED; &#xC778; &#xC0B0;&#xD654; Gli &#xB179;&#xC74C; &#xBC29;&#xC1A1; &#xC694;&#xC778;&#xACFC; Hh &#xC2DC;&#xADF8;&#xB110;&#xB9C1;&#xC758; &#xC548;&#xC815;&#xC131;&#xC5D0; &#xC911;&#xC694; &#xD55C; &#xC601;&#xD5A5;&#xC744;&#xAC00;&#xC9C0;&#xACE0;.

B56 &#xD3EC;&#xD568; &#xB41C; &#xB2E8;&#xBC31;&#xC9C8; &#xC778;&#xC0B0; &#xAC00;&#xC218;&#xBD84;&#xD574; &#xD6A8;&#xC18C; 2As &#xBC0F; &#xCE74; &#xC138; &#xC778; &#xB2C8; 2&#xC758; &#xB300;&#xB9BD; &#xD589;&#xB3D9; &#xC778; &#xC0B0;&#xD654; Daz &#xC0C1;&#xD638; &#xC791;&#xC6A9; &#xB2E8;&#xBC31;&#xC9C8; 1&#xC758; Gli &#xD68C;&#xC804;&#xC728; &#xAE30;&#xB2A5; &#xC81C;&#xC5B4;&#xD569;&#xB2C8;&#xB2E4;.

Vertebrate axis specification is an evolutionarily conserved developmental process that relies on asymmetric activation of Wnt signaling and subsequent organizer formation on the future dorsal side of the embryo. Although roles of Wnt signaling during organizer formation have been studied extensively, it is unclear how the Wnt pathway is asymmetrically activated. In Xenopus and zebrafish, the Wnt pathway is triggered by dorsal determinants, which are translocated from the vegetal pole to the future dorsal side of the embryo shortly after fertilization. The transport of dorsal determinants requires a unique microtubule network formed in the vegetal cortex shortly after fertilization. However, molecular mechanisms governing the formation of vegetal cortical microtubule arrays are not fully understood. Here we report that Dead-End 1 (Dnd1), an RNA-binding protein required for primordial germ cell development during later stages of embryogenesis, is essential for Xenopus axis specification. We show that knockdown of maternal Dnd1 specifically interferes with the formation of vegetal cortical microtubules. This, in turn, impairs translocation of dorsal determinants, the initiation of Wnt signaling, organizer formation, and ultimately results in ventralized embryos. Furthermore, we found that Dnd1 binds to a uridine-rich sequence in the 3'-UTR of trim36, a vegetally localized maternal RNA essential for vegetal cortical microtubule assembly. Dnd1 anchors trim36 to the vegetal cortex in the egg, promoting high concentrations of Trim36 protein there. Our work thus demonstrates a novel and surprising function for Dnd1 during early development and provides an important link between Dnd1, mRNA localization, the microtubule cytoskeleton and axis specification.

Maternal Dead-End1 is required for vegetal cortical microtubule assembly during Xenopus axis specification.

Recent genome-wide association studies reveal that the FAM13A gene is associated with human lung function and a variety of lung diseases, including chronic obstructive pulmonary disease, asthma, lung cancer, and pulmonary fibrosis. The biological functions of Fam13a, however, have not been studied. In an effort to identify novel substrates of B56-containing PP2As, we found that B56-containing PP2As and Akt act antagonistically to control reversible phosphorylation of Fam13a on Ser-322. We show that Ser-322 phosphorylation acts as a molecular switch to control the subcellular distribution of Fam13a. Fam13a shuttles between the nucleus and cytoplasm. When Ser-322 is phosphorylated by Akt, the binding between Fam13a and 14-3-3 is enhanced, leading to cytoplasmic sequestration of Fam13a. B56-containing PP2As dephosphorylate phospho-Ser-322 and promote nuclear localization of Fam13a. We generated Fam13a-knockout mice. Fam13a-mutant mice are viable and healthy, indicating that Fam13a is dispensable for embryonic development and physiological functions in adult animals. Intriguingly, Fam13a has the ability to activate the Wnt pathway. Although Wnt signaling remains largely normal in Fam13a-knockout lungs, depletion of Fam13a in human lung cancer cells causes an obvious reduction in Wnt signaling activity. Our work provides important clues to elucidating the mechanism by which Fam13a may contribute to human lung diseases.

Regulation of nuclear-cytoplasmic shuttling and function of Family with sequence similarity 13, member A (Fam13a), by B56-containing PP2As and Akt.

Hedgehog (Hh) signaling is fundamentally important for development and adult tissue homeostasis. It is well established that in vertebrates Sufu directly binds and inhibits Gli proteins, the downstream mediators of Hh signaling. However, it is unclear how the inhibitory function of Sufu towards Gli is regulated. Here we report that the Rusc family of proteins, the biological functions of which are poorly understood, form a heterotrimeric complex with Sufu and Gli. Upon Hh signaling, Rusc is displaced from this complex, followed by dissociation of Gli from Sufu. In mammalian fibroblast cells, knockdown of Rusc2 potentiates Hh signaling by accelerating signaling-induced dissociation of the Sufu-Gli protein complexes. In Xenopus embryos, knockdown of Rusc1 or overexpression of a dominant-negative Rusc enhances Hh signaling during eye development, leading to severe eye defects. Our study thus uncovers a novel regulatory mechanism controlling the response of cells to Hh signaling in vertebrates.

Members of the Rusc protein family interact with Sufu and inhibit vertebrate Hedgehog signaling.

A limited number of signaling pathways are repeatedly used to regulate a wide variety of processes during development and differentiation. The lack of tools to manipulate signaling pathways dynamically in space and time has been a major technical challenge for biologists. Optogenetic techniques, which utilize light to control protein functions in a reversible fashion, hold promise for modulating intracellular signaling networks with high spatial and temporal resolution. Applications of optogenetics in multicellular organisms, however, have not been widely reported. Here, we create an optimized bicistronic optogenetic system using Arabidopsis thaliana cryptochrome 2 (CRY2) protein and the N-terminal domain of cryptochrome-interacting basic-helix-loop-helix (CIBN). In a proof-of-principle study, we develop an optogenetic Raf kinase that allows reversible light-controlled activation of the Raf/MEK/ERK signaling cascade. In PC12 cells, this system significantly improves light-induced cell differentiation compared with co-transfection. When applied to Xenopus embryos, this system enables blue light-dependent reversible Raf activation at any desired developmental stage in specific cell lineages. Our system offers a powerful optogenetic tool suitable for manipulation of signaling pathways with high spatial and temporal resolution in a wide range of experimental settings.

Reversible optogenetic control of kinase activity during differentiation and embryonic development.

17β-estradiol is a potent sex hormone synthesized primarily by gonads in females and males that regulates development and function of the reproductive system. Recent studies show that 17β-estradiol is locally synthesized in nonreproductive tissues and regulates a myriad of events, including local inflammatory responses. In this study, we report that mesenteric lymph nodes (mLNs) and Peyer's patches (Pps) are novel sites of de novo synthesis of 17β-estradiol. These secondary lymphoid organs are located within or close to the gastrointestinal tract, contain leukocytes, and function at the forefront of immune surveillance. 17β-estradiol synthesis was initially identified using a transgenic mouse with red fluorescent protein coexpressed in cells that express aromatase, the enzyme responsible for 17β-estradiol synthesis. Subsequent immunohistochemistry and tissue culture experiments revealed that aromatase expression was localized to high endothelial venules of these lymphoid organs, and these high endothelial venule cells synthesized 17β-estradiol when isolated and cultured in vitro. Both mLNs and Pps contained 17β-estradiol with concentrations that were significantly higher than those of peripheral blood. Furthermore, the total amount of 17β-estradiol in these organs exceeded that of the gonads. Mice lacking either aromatase or estrogen receptor-β had hypertrophic Pps and mLNs with more leukocytes than their wild-type littermates, demonstrating a role for 17β-estradiol in leukocyte regulation. Importantly, we did not observe any sex-dependent differences in aromatase expression, 17β-estradiol content, or steroidogenic capacity in these lymphoid organs.

Estradiol Synthesis in Gut-Associated Lymphoid Tissue: Leukocyte Regulation by a Sexually Monomorphic System.

The intestinal epithelium plays a critical role in host-microbe homeostasis by sensing gut microbes and subsequently initiating proper immune responses. During the neonatal stage, the intestinal epithelium is under immune repression, allowing the transition for newborns from a relatively sterile intra-uterine environment to one that is rich in foreign antigens. The mechanism underlying such immune repression remains largely unclear, but involves downregulation of IRAK1 (interleukin-1 receptor-associated kinase), an essential component of toll-like receptor-mediated NF-κB signaling. We report here that heterogeneous nuclear ribonucleoprotein I (hnRNPI), an RNA binding protein, is essential for regulating neonatal immune adaptation. We generated a mouse model in which hnRNPI is ablated specifically in the intestinal epithelial cells, and characterized intestinal defects in the knockout mice. We found that loss of hnRNPI function in mouse intestinal epithelial cells results in early onset of spontaneous colitis followed by development of invasive colorectal cancer. Strikingly, the epithelium-specific hnRNPI knockout neonates contain aberrantly high IRAK1 protein levels in the colons and fail to develop immune tolerance to environmental microbes. Our results demonstrate that hnRNPI plays a critical role in establishing neonatal immune adaptation and preventing colitis and colorectal cancer.

Article	Year
Repression through a distal TCF-3 binding site restricts Xenopus myf-5 expression in gastrula mesoderm. Mechanisms of development\| PubMed ID: 12049769	2002
PP2A:B56epsilon is required for eye induction and eye field separation. Developmental biology\| PubMed ID: 17074314	2007
The 48-kDa alternative translation isoform of PP2A:B56epsilon is required for Wnt signaling during midbrain-hindbrain boundary formation. The Journal of biological chemistry\| PubMed ID: 19129191	2009
Heme metabolism enzymes are dynamically expressed during Xenopus embryonic development. Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al\| PubMed ID: 19181189	2008
hnRNP I inhibits Notch signaling and regulates intestinal epithelial homeostasis in the zebrafish. PLoS genetics\| PubMed ID: 19197356	2009
hnRNP I is required to generate the Ca2+ signal that causes egg activation in zebrafish. Development (Cambridge, England)\| PubMed ID: 19666827	2009
DUX4, a candidate gene for facioscapulohumeral muscular dystrophy, causes p53-dependent myopathy in vivo. Annals of neurology\| PubMed ID: 21446026	2011
The antagonistic action of B56-containing protein phosphatase 2As and casein kinase 2 controls the phosphorylation and Gli turnover function of Daz interacting protein 1. The Journal of biological chemistry\| PubMed ID: 21878643	2011
Maternal Dead-End1 is required for vegetal cortical microtubule assembly during Xenopus axis specification. Development (Cambridge, England)\| PubMed ID: 23615278	2013
Regulation of nuclear-cytoplasmic shuttling and function of Family with sequence similarity 13, member A (Fam13a), by B56-containing PP2As and Akt. Molecular biology of the cell\| PubMed ID: 25609086	2015
Members of the Rusc protein family interact with Sufu and inhibit vertebrate Hedgehog signaling. Development (Cambridge, England)\| PubMed ID: 27633991	2016
Reversible optogenetic control of kinase activity during differentiation and embryonic development. Development (Cambridge, England)\| PubMed ID: 27697903	2016
Estradiol Synthesis in Gut-Associated Lymphoid Tissue: Leukocyte Regulation by a Sexually Monomorphic System. Endocrinology\| PubMed ID: 27779914	2016
hnRNP I regulates neonatal immune adaptation and prevents colitis and colorectal cancer. PLoS genetics\| PubMed ID: 28296893	2017

Wenyan Mei

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JoVE Journal Publications

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