yeast-luminometric-xenopus-oocyte-electrophysiological-examinations

Yeast Luminometric and <em>Xenopus</em> Oocyte Electrophysiological Examinations of the Molecular Mechanosensitivity of TRPV4

TRPV4 (Transient Receptor Potentials, vanilloid family, type 4) is widely expressed in vertebrate tissues and is activated by several stimuli, including ...

University of Wisconsin – Madison

Transient receptor potential (TRP) channels are first elements in sensing chemicals, heat, and force and are widespread among protists and fungi as well as animals. Despite their importance, the arrangement and roles of the amino acids that constitute the TRP channel gate are unknown. The yeast TRPY1 is activated in vivo by osmotically induced vacuolar membrane deformation and by cytoplasmic Ca(2+). After a random mutagenesis, we isolated TRPY1 mutants that responded more strongly to mild osmotic upshocks. One such gain-of-function mutant has a Y458H substitution at the C terminus of the predicted sixth transmembrane helix. Direct patch-clamp examination of vacuolar membranes showed that Y458H channels were already active with little stimulus and showed marked flickers between the open and intraburst closed states. They remained responsive to membrane stretch force and to Ca(2+), indicating primary defects in the gate region but not in the sensing of gating principles. None of the other 18 amino acid replacements engineered here showed normal channel kinetics except the two aromatic substitutions, Y458F and Y458W. The Y458 of TRPY1 has its aromatic counterpart in mammalian TRPM. Furthermore, conserved aromatics one alpha-helical turn downstream from this point are also found in animal TRPC, TRPN, TRPP, and TRPML, suggesting that gate anchoring with aromatics may be common among many TRP channels. The possible roles of aromatics at the end of the sixth transmembrane helix are discussed.

Yeast screens show aromatic residues at the end of the sixth helix anchor transient receptor potential channel gate.

Transient receptor potential (TRP) channels found in animals, protists, and fungi are primary chemo-, thermo-, or mechanosensors. Current research emphasizes the characteristics of individual channels in each animal TRP subfamily but not the mechanisms common across subfamilies. A forward genetic screen of the TrpY1, the yeast TRP channel, recovered gain-of-function (GOF) mutations with phenotype in vivo and in vitro. Single-channel patch-clamp analyses of these GOF-mutant channels show prominent aberrations in open probability and channel kinetics. These mutations revealed functionally important aromatic amino acid residues in four locations: at the intracellular end of the fifth transmembrane helix (TM5), at both ends of TM6, and at the immediate extension of TM6. These aromatics have counterparts in most TRP subfamilies. The one in TM5 (F380L) aligns precisely with an exceptional Drosophila mutant allele (F550I) that causes constitutive activity in the canonical TRP channel, resulting in rapid and severe retinal degeneration beyond mere loss of phototaxis. Thus, this phenylalanine maintains the balance of various functional states (conformations) of a channel for insect phototransduction as well as one for fungal mechanotransduction. This residue is among a small cluster of phenylalanines found in all known subfamilies of TRP channels. This unique case illustrates that GOF mutations can reveal structure-function principles that can be generalized across different TRP subfamilies. It appears that the conserved aromatics in the four locations have conserved functions in most TRP channels. The possible mechanistic roles of these aromatics and the further use of yeast genetics to dissect TRP channels are discussed.

Yeast gain-of-function mutations reveal structure-function relationships conserved among different subfamilies of transient receptor potential channels.

In yeast, osmotic upshock causes a release of vacuolar Ca(2+) through the mechanosensitive transient receptor potential channel, Yvc1. We screened the collection of 4810 yeast gene deletants twice for alterations in this response in an attempt to find elements that regulate the amount of vacuolar Ca(2+) or the Yvc1 channel. Severe overresponders and underresponders to upshock were further scrutinized for their calcium content with (45)Ca and their Yvc1 electrophysiological activities under patch-clamp. The severe underresponders have lower calcium content but no change in Yvc1 activity. The strong overresponders, most of which are deleted of genes involved in cell wall metabolism, have higher calcium content. Wall mutations are known to up-regulate Ca(2+)-calcineurin-dependent genes. It appears that stress on the cell wall induces Ca(2+) accumulation, adaptively anticipating the need in defense or repair against future stress, including osmotic stress.

A genome-wide survey suggests an osmoprotective role for vacuolar Ca2+ release in cell wall-compromised yeast.

The ability to sense mechanical and osmotic stimuli is vital to all organisms from mammals to bacteria. Members of the transient receptor potential (TRP) ion-channel family have attracted intense attention for their involvement in mechanosensation. The yeast homologue TRPY1 can clearly be activated by hypertonic shock in vivo and by stretch force under patch clamp. Like its animal counterparts, TRPY1 is polymodal, being gated by membrane stretch force and by cytoplasmic Ca(2+). Here, we investigated how these two gating principles interact. We found that stretch force can induce some channel activation without cytoplasmic Ca(2+). Tens of micromolar Ca(2+) greatly enhance the observed force-induced activities, with open probabilities following well the Boltzmann distribution, in which the two gating energies are summed as exponents. To map this formalism to structures, we found Ca(2+)-binding proteins such as calmodulin or calcineurin to be unnecessary. However, removing a dense cluster of negative charges in the C-terminal cytoplasmic domain of TRPY1 greatly diminishes the Ca(2+) activation as well as its influence on force activation. We also found a strategic point upstream of this charge cluster, at which insertion of amino acids weakens Ca(2+) activation considerably but leaves the mechanosensitivity nearly intact. These results led to a structure-function model in which Ca(2+) binding to the cytoplasmic domain and stretching of the membrane-embedded domain both generate gating force, reaching the gate in parallel.

Mechanical force and cytoplasmic Ca(2+) activate yeast TRPY1 in parallel.

Mechanosensitive (MS) ion channels likely underlie myriad force-sensing processes, from basic osmotic regulation to specified sensations of animal hearing and touch. Albeit important, the molecular identities of many eukaryotic MS channels remain elusive, let alone their working mechanisms. This is in stark contrast to our advanced knowledge on voltage- or ligand-sensitive channels. Several members of transient receptor potential (TRP) ion channel family have been implicated to function in mechanosensation and are recognized as promising candidate MS channels. The yeast TRP homolog, TRPY1, is clearly a first-line force transducer. It can be activated by hypertonic shock in vivo and by membrane stretch force in excised patches under patch clamp, making it a useful model for understanding TRP channel mechanosensitivity in general. TRPY1 offers two additional research advantages: (1) It has a large ( approximately 300 pS) unitary conductance and therefore a favorable S/N ratio. (2) Budding yeast allows convenient and efficient genetic and molecular manipulations. In this review, we focus on the current research of TRPY1 and discuss its prospect. We also describe the use of yeast as a system to express and characterize animal TRP channels.

The use of yeast to understand TRP-channel mechanosensitivity.

TRPV4 is a polymodal cation channel gain-of-function (GOF) allele which causes skeletal dysplasia in humans. To better understand its gating, we screened for additional GOF alleles based on their ability to block yeast proliferation. Repeatedly, only a limited number of such growth-blocking mutations were isolated. Expressed in oocytes, wild-type channels can be strongly activated by either hypotonicity or exposure to the potent agonist 4alphaPDD, although the GOF channels behaved as if they were fully prestimulated as well as lacking a previously uncharacterized voltage-dependent inactivation. Five of six mutations occurred at or near the inner ends of the predicted core helices, giving further direct evidence that this region indeed forms the main intracellular gate in TRP channels. Surprisingly, both wild-type channels as well as these GOF channels maintain strong steady-state outward rectification that is not due to a Ca(2+) block, as has been proposed elsewhere. We conclude that TRPV4 contains an additional voltage-dependent gating mechanism in series with the main intracellular gate.

Forward genetic analysis reveals multiple gating mechanisms of TRPV4.

Whether animal ion channels functioning as mechanosensors are directly activated by stretch force or indirectly by ligands produced by the stretch is a crucial question. TRPV4, a key molecular model, can be activated by hypotonicity, but the mechanism of activation is unclear. One model has this channel being activated by a downstream product of phospholipase A(2), relegating mechanosensitivity to the enzymes or their regulators. We expressed rat TRPV4 in Xenopus oocytes and repeatedly examined >200 excised patches bathed in a simple buffer. We found that TRPV4 can be activated by tens of mm Hg pipette suctions with open probability rising with suction even in the presence of relevant enzyme inhibitors. Mechanosensitivity of TRPV4 provides the simplest explanation of its various force-related physiological roles, one of which is in the sensing of weight load during bone development. Gain-of-function mutants cause heritable skeletal dysplasias in human. We therefore examined the brachyolmia-causing R616Q gain-of-function channel and found increased whole-cell current densities compared with wild-type channels. Single-channel analysis revealed that R616Q channels maintain mechanosensitivity but have greater constitutive activity and no change in unitary conductance or rectification.

Wild-type and brachyolmia-causing mutant TRPV4 channels respond directly to stretch force.

To date, dozens of melanoma-associated antigens (MAGE) have been identified. Based on the differences in tissue-specific gene expression and gene structure, the MAGE family has been divided into two big subfamilies: MAGE-I and MAGE-II. MAGE-I genes were identified as a group of highly attractive targets for cancer immunotherapy because of their wide expression in a variety of malignant tumors but silent in normal adult cells except germ-line cells lacking human leukocyte antigen (HLA) expression. However, little is known regarding the functions of MAGE family members in cell activities. In this review, we briefly described the classification of MAGE family members and their expression pattern in cancer. We also summarized the mechanism of MAGE activation and the functions of MAGE family members in cell cycle progression and apoptosis. We also discussed what is known of immunotherapy targeting MAGE family.

Melanoma-associated antigen genes - an update.

The melonoma-associated antigens family A (MAGE-A) belongs to cancer/testis antigens (CTA) that are expressed in a wide variety of malignant tumors but not in normal adult tissues except for testis. Interestingly, germ cells do not express MHC class I antigen, implying that these gene products should be ideal targets for cancer immunotherapy. The strict tumor-specific expression of MAGE-As has led to several immunotherapeutic trials targeting some of these proteins. In this review, we briefly described the expression and activation mechanism of MAGE-As in cancer. We also summarized the biological functions of MAGE-As in cell progress and the progress of the cancer immunotherapy targeting MAGE-A family.

MAGE-A family: attractive targets for cancer immunotherapy.

Melanoma-associated antigens-A (MAGE-A) family is a group of well-characterized cancer/testis antigens (CTA), because they are strictly tumor specific and are shared by many kinds of tumors. However, the expression pattern of MAGE-A10 and MAGE-A11 in breast cancer patients is still unclear. The purpose of our study is to investigate the expression pattern and prognostic significance of MAGE-A10 and MAGE-A11 in breast cancer patients.

Expressions of MAGE-A10 and MAGE-A11 in breast cancers and their prognostic significance: a retrospective clinical study.

&#x6797;&#x57DF;&#x552F;&#x4E00;&#x86CB;&#x767D;&#x8D28; (&#x6539;&#x6027;&#x6D3B;&#x751F;&#x7269;&#x4F53;&#xFF09; &#x7EC4;&#x6210;&#x7684;&#x56DB;&#x540D;&#x6210;&#x5458; (LMO1 LMO4) &#x662F;&#x4E00;&#x5BB6;&#x4EBA;&#x7684;&#x7279;&#x70B9;&#x662F;&#x72EC;&#x5360;&#x7684;&#x4E24;&#x4E2A;&#x4E32;&#x8054;&#x6797;&#x57DF;&#x548C;&#x4EFB;&#x4F55;&#x5176;&#x4ED6;&#x529F;&#x80FD;&#x7684;&#x57DF;&#x5B58;&#x5728;&#x7684;&#x6838;&#x8F6C;&#x5F55; coregulators&#x3002;&#x4ED6;&#x4EEC;&#x901A;&#x8FC7;&#x8FD0;&#x4F5C;&quot;&#x94FE;&#x63A5;&#x5668;&quot;&#x6216;&quot;&#x811A;&#x624B;&#x67B6;&quot;&#x86CB;&#x767D;&#x8D28;&#x6839;&#x636E;&#x5176;&#x6797;&#x57DF;&#x4F5C;&#x4E3A;&#x8C03;&#x8282;&#x57FA;&#x56E0;&#x8F6C;&#x5F55;&#xFF0C;&#x6D89;&#x53CA;&#x51E0;&#x4E2A; DNA &#x7ED3;&#x5408;&#x56E0;&#x5B50;&#x548C;&#x8F6C;&#x5F55;&#x8C03;&#x8282;&#x86CB;&#x767D;&#x7684; multiprotein &#x914D;&#x5408;&#x7269;&#x7684;&#x5F62;&#x6210;&#x3002;&#x5728;&#x672C;&#x7814;&#x7A76;&#x4E2D;&#xFF0C;&#x6211;&#x4EEC;&#x8BD5;&#x56FE;&#x627E;&#x5230; p53 &#x548C; LMO4 &#x548C; LMO4 &#x5BF9; p53 &#x4ECB;&#x5BFC;&#x7684;&#x4E73;&#x817A;&#x764C;&#x7EC6;&#x80DE;&#x589E;&#x6B96;&#x6291;&#x5236;&#x4F5C;&#x7528;&#x5F71;&#x54CD;&#x7684;&#x7269;&#x7406;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x3002;

LMO4 &#x6291;&#x5236; p53 &#x4ECB;&#x5BFC;&#x901A;&#x8FC7;&#x76F8;&#x4E92;&#x4F5C;&#x7528; p53 &#x4E73;&#x817A;&#x764C;&#x7EC6;&#x80DE;&#x589E;&#x6B96;&#x7684;&#x6291;&#x5236;&#x3002;

The LIM domain only proteins (LMOs) which consist of four members (LMO1-LMO4) are a family of nuclear transcription coregulators that are characterized by the exclusive presence of two tandem LIM domains and no other functional domains. They regulate gene transcription by functioning as "linker" or "scaffolding" proteins by virtue of their LIM domains and are involved in the formation of multiprotein complexes with several DNA-binding factors and transcriptional regulatory proteins. In the present study, we tried to find the physical interaction between p53 and LMO4, and the effect of LMO4 on p53-mediated proliferative inhibition of breast cancer cells.

LMO4 inhibits p53-mediated proliferative inhibition of breast cancer cells through interacting p53.

Les LIM domaine seules prot&eacute;ines (OVM) qui sont compos&eacute;es de quatre membres (LMO1-LMO4) sont une famille de cor&eacute;gulateurs de transcription nucl&eacute;aire qui se caract&eacute;risent par la pr&eacute;sence exclusive de deux domaines LIM en tandem et aucun des autres domaines fonctionnels. Ils r&eacute;gulent la transcription du g&egrave;ne en fonctionnant comme &laquo; linker &raquo; ou les prot&eacute;ines &laquo; &eacute;chafaudage &raquo; en vertu de leurs domaines LIM et sont impliqu&eacute;s dans la formation de complexes multiprot&eacute;iques avec plusieurs facteurs de liaison &agrave; l&#39;ADN et les prot&eacute;ines de r&eacute;gulation transcriptionnelles. Dans la pr&eacute;sente &eacute;tude, nous avons essay&eacute; de trouver l&#39;interaction physique entre l&#39;effet du LMO4 sur l&#39;inhibition prolif&eacute;rative de cellules canc&eacute;reuses du sein par une p53 p53 et LMO4.

LMO4 inhibe l&#39;inhibition prolif&eacute;rative p53-par des cellules canc&eacute;reuses du sein gr&acirc;ce &agrave; l&#39;interaction de p53.

LIM Dom&auml;ne nur Proteine (LVO) besteht aus vier Mitgliedern (LMO1-LMO4) sind eine Familie der nuklearen Transkription Coregulators, die durch das ausschlie&szlig;liche Vorhandensein von zwei Tandem LIM und keine andere funktionellen Dom&auml;nen gekennzeichnet sind. Sie gen-Transkription zu regulieren, indem Sie als &quot;Linker&quot; oder &quot;Ger&uuml;st&quot; Proteine aufgrund ihrer LIM-Domains funktionieren und sind an der Bildung der multiprotein komplexe mit mehreren DNA-bindenden Faktoren und transkriptionelle regulatorischen Proteine beteiligt. In der vorliegenden Studie haben wir versucht, die k&ouml;rperliche Interaktion zwischen p53 LMO4 und die Wirkung des LMO4 auf p53-vermittelte proliferative Hemmung der Brustkrebs-Zellen zu finden.

LMO4 hemmt p53-vermittelte proliferative Hemmung der Brustkrebs-Zellen durch Interaktion p53.

LIM dominio sola proteine (OVM) che consistono di quattro membri (LMO1-LMO4) sono una famiglia di coregulators della trascrizione nucleare che sono caratterizzati dalla presenza esclusiva di due tandem LIM domini e nessun altro funzionale. Essi regolano la trascrizione del gene di funzionamento come &quot;linker&quot; o proteine &quot;impalcatura&quot; in virt&ugrave; della loro domini LIM e sono coinvolti nella formazione di complessi multiproteici con diversi fattori DNA-binding e transcriptional proteine regolatorie. Nel presente studio, abbiamo cercato di trovare l&#39;interazione fisica tra p53 e LMO4 e l&#39;effetto di LMO4 sull&#39;inibizione proliferativa p53-mediata delle cellule di cancro al seno.

LMO4 inibisce la inibizione proliferativa p53-mediata delle cellule di cancro al seno attraverso interazione p53.

4 &#x4EBA;&#x306E;&#x30E1;&#x30F3;&#x30D0;&#x30FC; (LMO1 LMO4) &#x3067;&#x69CB;&#x6210;&#x3055;&#x308C;&#x308B;&#x3001;LIM &#x30C9;&#x30E1;&#x30A4;&#x30F3;&#x306E;&#x307F;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA; (Lmo) 2 &#x3064;&#x306E;&#x30BF;&#x30F3;&#x30C7;&#x30E0; LIM &#x30C9;&#x30E1;&#x30A4;&#x30F3;&#x3068;&#x306A;&#x3044;&#x4ED6;&#x306E;&#x6A5F;&#x80FD;&#x30C9;&#x30E1;&#x30A4;&#x30F3;&#x306E;&#x6392;&#x4ED6;&#x7684;&#x306A;&#x5B58;&#x5728;&#x306B;&#x3088;&#x3063;&#x3066;&#x7279;&#x5FB4;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x308B;&#x6838;&#x306E;&#x8EE2;&#x5199;&#x30B3;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30BF;&#x30FC;&#x306E;&#x5BB6;&#x65CF;&#x3067;&#x3059;&#x3002;&#x5F7C;&#x3089;&#x306F;&#x3001;&#x300C;&#x30EA;&#x30F3;&#x30AB;&#x30FC;&#x300D;&#x307E;&#x305F;&#x306F;&#x305D;&#x306E; LIM &#x30C9;&#x30E1;&#x30A4;&#x30F3;&#x306E;&#x300C;&#x8DB3;&#x5834;&#x300D;&#x86CB;&#x767D;&#x8CEA;&#x3068;&#x3057;&#x3066;&#x6A5F;&#x80FD;&#x306B;&#x3088;&#x3063;&#x3066;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x8EE2;&#x5199;&#x3092;&#x8ABF;&#x7BC0;&#x3059;&#x308B;&#x3001;&#x3044;&#x304F;&#x3064;&#x304B;&#x306E; DNA &#x7D50;&#x5408;&#x56E0;&#x5B50;&#x3068;&#x8EE2;&#x5199;&#x8ABF;&#x7BC0;&#x86CB;&#x767D;&#x8CEA;&#x3067;&#x591A;&#x86CB;&#x767D;&#x8CEA;&#x8907;&#x5408;&#x4F53;&#x5F62;&#x6210;&#x306B;&#x95A2;&#x4E0E;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3044;&#x307E;&#x3059;&#x3002;&#x672C;&#x7814;&#x7A76;&#x3067;&#x306F;&#x3001;&#x6211;&#x3005; &#x3068; p53 &#x3068; LMO4&#x3001;&#x4E73;&#x304C;&#x3093;&#x7D30;&#x80DE;&#x306E;&#x5897;&#x6B96;&#x963B;&#x5BB3;&#x306E; p53 &#x3092;&#x4ECB;&#x3059;&#x308B;&#x306B;&#x53CA;&#x307C;&#x3059; LMO4 &#x306E;&#x7269;&#x7406;&#x7684;&#x306A;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x3092;&#x898B;&#x3064;&#x3051;&#x308B;&#x3053;&#x3068;&#x3092;&#x8A66;&#x307F;&#x305F;&#x3002;

LMO4 p53 &#x3092;&#x4ECB;&#x3059;&#x308B;&#x3092;&#x4ECB;&#x3057;&#x3066;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x3059;&#x308B; p53 &#x306E;&#x4E73;&#x623F;&#x306E;&#x304C;&#x3093;&#x7D30;&#x80DE;&#x5897;&#x6B96;&#x963B;&#x5BB3;&#x3092;&#x6291;&#x5236;&#x3057;&#x307E;&#x3059;&#x3002;

&#xC784; &#xB3C4;&#xBA54;&#xC778;&#xB9CC; &#xB2E8;&#xBC31;&#xC9C8; (LMOs) 4 &#xBA85; (LMO1 LMO4)&#xC73C;&#xB85C; &#xAD6C;&#xC131; &#xB41C;&#xB294; &#xB450; &#xC9C1;&#xB82C; &#xC2DD; &#xC784; &#xB3C4;&#xBA54;&#xC778;&#xACFC; &#xB2E4;&#xB978; &#xAE30;&#xB2A5; &#xB3C4;&#xBA54;&#xC778;&#xC758; &#xB3C5;&#xC810;&#xC801;&#xC778; &#xC874;&#xC7AC;&#xB97C; &#xD2B9;&#xC9D5;&#xC73C;&#xB85C;&#xD558;&#xB294; &#xD575; &#xC804;&#xC0AC; coregulators&#xC758; &#xAC00;&#xC871;. &#xADF8;&#xB4E4;&#xC740; &quot;&#xB9C1;&#xCEE4;&quot; &#xB610;&#xB294; &quot;&#xBE44; &#xACC4;&quot; &#xB2E8;&#xBC31;&#xC9C8; &#xC784; &#xB3C4;&#xBA54;&#xC778;&#xC758; &#xB355;&#xC73C;&#xB85C; &#xC791;&#xC6A9; &#xD558; &#xC5EC; &#xC720;&#xC804;&#xC790; &#xC804;&#xC0AC;&#xB97C; &#xC870;&#xC808; &#xD558; &#xACE0; &#xC5EC;&#xB7EC; DNA &#xBC14;&#xC778;&#xB529; &#xC694;&#xC18C;&#xC640; transcriptional &#xADDC;&#xC81C; &#xB2E8;&#xBC31;&#xC9C8; multiprotein &#xB2E8;&#xC9C0;&#xC758; &#xD615;&#xC131;&#xC5D0; &#xAD00;&#xC5EC; &#xD558;&#xB294;. &#xD604;&#xC7AC;&#xC758; &#xC5F0;&#xAD6C;&#xC5D0;&#xC11C; p53 &#xBC0F; LMO4, &#xC720;&#xBC29;&#xC554; &#xC138;&#xD3EC;&#xC758; &#xC99D;&#xC2DD; &#xC5B5;&#xC81C; p53 &#xC911;&#xC7AC;&#xC5D0; lmo4&#xC758; &#xD6A8;&#xACFC; &#xC0AC;&#xC774;&#xC758; &#xC2E4;&#xC81C; &#xC0C1;&#xD638; &#xC791;&#xC6A9;&#xC744; &#xCC3E;&#xC544; &#xBCF4;&#xC558;&#xC2B5;&#xB2C8;&#xB2E4;.

LMO4 &#xC0C1;&#xD638; &#xC791;&#xC6A9; p53 &#xD1B5;&#xD574; &#xC720;&#xBC29;&#xC554; &#xC138;&#xD3EC;&#xC758; &#xC99D;&#xC2DD; &#xC5B5;&#xC81C;&#xB97C; p53 &#xC911;&#xC7AC;&#xB97C; &#xC5B5;&#xC81C; &#xD55C;&#xB2E4;.

LIM dominio s&oacute;lo prote&iacute;nas (OVM) que consisten en cuatro miembros (LMO1-LMO4) son una familia de coregulators de la transcripci&oacute;n nuclear que se caracterizan por la presencia exclusiva de dos dominios LIM t&aacute;ndem y no otros dominios funcionales. Regulan la transcripci&oacute;n gen&eacute;tica por funcionar como &quot;linker&quot; o prote&iacute;nas &quot;andamios&quot; en virtud de sus dominios LIM y participan en la formaci&oacute;n de Complejos Multiproteinas con varios factores de uni&oacute;n al ADN y prote&iacute;nas reguladoras transcripcionales. En el presente estudio, hemos intentado encontrar la interacci&oacute;n f&iacute;sica entre la p53 y LMO4 y el efecto del LMO4 sobre la inhibici&oacute;n proliferativa mediada por p53 de las c&eacute;lulas de c&aacute;ncer de mama.

LMO4 inhibe la inhibici&oacute;n proliferativa mediada por p53 de las c&eacute;lulas de c&aacute;ncer de mama a trav&eacute;s de la interacci&oacute;n de la p53.

Abstract Ovarian cancer is a leading cause of death among gynecologic malignancies. In this study, we reported the expression of melanoma-associated antigens A (MAGE-A) genes in peripheral blood from 80 patients with ovarian cancer and 30 healthy donors. MAGE-As expression was associated with the factors indicating poor prognosis. The expressions of MAGE-As and each individual MAGE-A genes were also associated with low overall survival of patients with ovarian cancer. Our results suggested MAGE-A genes may have the potential to be surveillance markers for the detection of circulating tumor cells and represent a poor prognosis for patients with ovarian cancer.

Multiple MAGE-A genes as surveillance marker for the detection of circulating tumor cells in patients with ovarian cancer.

The LIM-only subclass of LIM proteins is a family of nuclear transcription co-regulators that are characterized by the exclusive presence of two tandem LIM domains and no other functional domains. To date, four LIM-domain-only (LMO) proteins (LMO1-LMO4) have been identified. They regulate gene transcription by functioning as "linker" or "scaffolding" proteins with a remarkable potential to mediate protein-protein interactions. These proteins play important roles in cell fate determination, cell growth and differentiation, tissues patterning, and organ development. In this review, we briefly described the functions of LMO proteins in the organ development and diseases. We also summarized the interaction proteins of each LMO family member, which may contribute to elucidating the functions of these mysterious and important linker proteins.

Article	Year
Yeast screens show aromatic residues at the end of the sixth helix anchor transient receptor potential channel gate. Proceedings of the National Academy of Sciences of the United States of America\| PubMed ID: 17878311	2007
Yeast gain-of-function mutations reveal structure-function relationships conserved among different subfamilies of transient receptor potential channels. Proceedings of the National Academy of Sciences of the United States of America\| PubMed ID: 18042709	2007
A genome-wide survey suggests an osmoprotective role for vacuolar Ca2+ release in cell wall-compromised yeast. FASEB journal : official publication of the Federation of American Societies for Experimental Biology\| PubMed ID: 18323404	2008
Mechanical force and cytoplasmic Ca(2+) activate yeast TRPY1 in parallel. The Journal of membrane biology\| PubMed ID: 19219385	2009
The use of yeast to understand TRP-channel mechanosensitivity. Pflügers Archiv : European journal of physiology\| PubMed ID: 19462180	2009
Forward genetic analysis reveals multiple gating mechanisms of TRPV4. The Journal of biological chemistry\| PubMed ID: 20424166	2010
Wild-type and brachyolmia-causing mutant TRPV4 channels respond directly to stretch force. The Journal of biological chemistry\| PubMed ID: 20605796	2010
Melanoma-associated antigen genes - an update. Cancer letters\| PubMed ID: 21093980	2011
MAGE-A family: attractive targets for cancer immunotherapy. Vaccine\| PubMed ID: 21933694	2011
Expressions of MAGE-A10 and MAGE-A11 in breast cancers and their prognostic significance: a retrospective clinical study. Journal of cancer research and clinical oncology\| PubMed ID: 22198676	2012
LMO4 inhibits p53-mediated proliferative inhibition of breast cancer cells through interacting p53. Life sciences\| PubMed ID: 22906635	2012
Multiple MAGE-A genes as surveillance marker for the detection of circulating tumor cells in patients with ovarian cancer. Biomarkers : biochemical indicators of exposure, response, and susceptibility to chemicals\| PubMed ID: 24320162	2014
LIM-domain-only proteins: multifunctional nuclear transcription coregulators that interacts with diverse proteins. Molecular biology reports\| PubMed ID: 24379077	2014

Xinliang Zhou

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