a-comparative-approach-to-characterize-landscape-host-pathogen

A Comparative Approach to Characterize the Landscape of Host-Pathogen Protein-Protein Interactions

Significant efforts were gathered to generate  large-scale comprehensive protein-protein interaction network maps. This is  instrumental to understand the ...

Institut Pasteur 

Mechanisms of cellular transformation associated with human papillomavirus type 5 (HPV5), which is responsible for skin carcinomas in epidermodysplasia verruciformis (EV) patients, are poorly understood. Using a yeast two-hybrid screening and molecular and cellular biology experiments, we found that HPV5 oncoprotein E6 interacts with SMAD3, a key component in the transforming growth factor beta1 (TGF-beta1) signaling pathway. HPV5 E6 inhibits SMAD3 transactivation by destabilizing the SMAD3/SMAD4 complex and inducing the degradation of both proteins. Interestingly, the E6 protein of nononcogenic EV HPV9 failed to interact with SMAD3, suggesting that downregulation of the TGF-beta1 signaling pathway could be a determinant in HPV5 skin carcinogenesis.

Human papillomavirus type 5 E6 oncoprotein represses the transforming growth factor beta signaling pathway by binding to SMAD3.

M&eacute;canismes de transformation cellulaire associ&eacute; &agrave; virus du papillome humain type 5 (HPV5), qui est responsable des carcinomes de la peau chez les patients d&#39;&eacute;pidermodysplasie verruciforme (EV), sont mal connus. &Agrave; l&#39;aide d&#39;une projection de deux-hybride de levure et des exp&eacute;riences de biologie mol&eacute;culaire et cellulaire, nous avons constat&eacute; que HPV5 oncoprot&eacute;ine E6 interagit avec SMAD3, un &eacute;l&eacute;ment cl&eacute; dans la beta1 de facteur de croissance transformant (TGF-beta1) voie de signalisation. HPV5 E6 inhibe SMAD3 transactivation en d&eacute;stabilisant le complexe SMAD3/SMAD4 et en induisant la d&eacute;gradation de ces deux prot&eacute;ines. Fait int&eacute;ressant, la prot&eacute;ine E6 de nononcogenic EV HPV9 n&#39;a pas pu interagir avec SMAD3, ce qui sugg&egrave;re que la diminution de la TGF-beta1 signalisation voie pourrait &ecirc;tre un facteur d&eacute;terminant dans la carcinogen&egrave;se de peau HPV5.

Virus du papillome humain type 5 E6 oncoprot&eacute;ine r&eacute;prime le transforming growth factor beta voie en se liant &agrave; SMAD3 de signalisation.

Meccanismi di trasformazione cellulare associato a papillomavirus umano di tipo 5 (HPV5), che &egrave; responsabile per i carcinomi della pelle nei pazienti di Epidermodisplasia verruciforme (EV), sono mal capiti. Utilizzando una proiezione di due-ibrido del lievito ed esperimenti di biologia molecolare e cellulare, abbiamo trovato che HPV5 oncoproteina dei tipo E6 interagisce con SMAD3, un componente chiave nella beta1 di fattore di crescita trasformante (TGF-beta1) signaling pathway. HPV5 E6 inibisce SMAD3 transactivation di destabilizzare il complesso SMAD3/SMAD4 e indurre la degradazione di entrambe le proteine. Curiosamente, la proteina E6 di HPV EV HPV9 non &egrave; riuscito a interagire con SMAD3, suggerendo che la downregulation del TGF-beta1 signaling pathway potrebbe essere un fattore determinante nella carcinogenesi pelle HPV5.

Human papillomavirus type 5 E6 oncoproteina dei tipo reprime il transforming growth factor beta signaling pathway legandosi a SMAD3.

To examine conjunctival papilloma and normal conjunctival tissue for the presence of human papillomavirus (HPV).

Human papillomavirus in normal conjunctival tissue and in conjunctival papilloma: types and frequencies in a large series.

D&#39;examiner le papillome conjonctival et tissu conjonctival normal pour la pr&eacute;sence du virus du papillome humain (VPH).

Virus du papillome humain dans le tissu conjonctival normal et en papillome conjonctival : types et fr&eacute;quences dans une grande s&eacute;rie.

Per esaminare il papilloma congiuntiva e tessuto congiuntiva normale per la presenza di papillomavirus umano (HPV).

Papillomavirus umano nel normale tessuto congiuntiva e nel papilloma congiuntiva: tipi e le frequenze in una serie di grandi dimensioni.

Epithelial tumours of the lacrimal sac are rare but important entities that may carry grave prognoses. In this study the prevalence and possible role of human papillomavirus (HPV) infection in epithelial tumours of the lacrimal sac were evaluated.

Human papillomavirus: cause of epithelial lacrimal sac neoplasia?

Les tumeurs &eacute;pith&eacute;liales du sac lacrymal sont rares mais importantes des entit&eacute;s qui peuvent porter des pronostics graves. Dans cette &eacute;tude, la pr&eacute;valence et le r&ocirc;le possible de l&#39;infection &agrave; papillomavirus humain (HPV) dans les tumeurs &eacute;pith&eacute;liales du sac lacrymal ont &eacute;t&eacute; &eacute;valu&eacute;es.

Virus du papillome humain : provoquer des n&eacute;oplasies &eacute;pith&eacute;liales sac lacrymal ?

Tumori epiteliali del sacco lacrimale sono entit&agrave; rara ma importante che pu&ograve; portare a prognosi grave. In questo studio sono state valutate la prevalenza e il possibile ruolo dell&#39;infezione da papillomavirus umano (HPV) nei tumori epiteliali del sacco lacrimale.

Papillomavirus umano: causa di neoplasie epiteliali sacco lacrimale?

We identified sequences from two distantly related papillomaviruses in genital warts from two Burmeister's porpoises, including a PV antigen-positive specimen, and characterized Phocoena spinipinnis papillomavirus type 1 (PsPV-1). The PsPV-1 genome comprises 7879 nt and presents unusual features. It lacks an E7, an E8 and a bona fide E5 open reading frame (ORF) and has a large E6 ORF. PsPV-1 L1 ORF showed the highest percentage of nucleotide identity (54-55 %) with human papillomavirus type 5, bovine papillomavirus type 3 (BPV-3) and Tursiops truncatus papillomavirus type 2 (TtPV-2). This warrants the classification of PsPV-1 as the prototype of the genus Omikronpapillomavirus. PsPV-1 clustered with TtPV-2 in the E6 and E1E2 phylogenetic trees and with TtPV-2 and BPV-3 in the L2L1 tree. This supports the hypothesis that PV evolution may not be monophyletic across all genes.

Genital warts in Burmeister's porpoises: characterization of Phocoena spinipinnis papillomavirus type 1 (PsPV-1) and evidence for a second, distantly related PsPV.

Nous avons identifi&eacute; les s&eacute;quences de deux papillomavirus lointainement connexes dans les verrues g&eacute;nitales de marsouin de Burmeister deux, dont un sp&eacute;cimen d&#39;antig&egrave;ne-positif de PV et caract&eacute;rise Phocoena spinipinnis papillomavirus de type 1 (PsPV-1). Le g&eacute;nome de PsPV-1 comprend 7879 nt et pr&eacute;sente des caract&eacute;ristiques inhabituelles. Il manque un E7, un E8 et un v&eacute;ritable cadre de lecture ouvert E5 (ORF) et a un grand ORF de E6. PsPV-1 L1 ORF a montr&eacute; le plus fort pourcentage d&#39;identit&eacute; de nucl&eacute;otides (54-55 %) avec le virus du papillome humain type 5, bovine papillomavirus de type 3 (VPB-3) et Tursiops truncatus papillomavirus de type 2 (TtPV-2). Cela garantit que la classification de PsPV-1 comme le prototype du genre Omikronpapillomavirus. PsPV-1 en cluster avec TtPV-2 dans les arbres phylog&eacute;n&eacute;tiques E6 et E1E2 et TtPV-2 et VPB-3 dans l&#39;arborescence de L2L1. Ceci appuie l&#39;hypoth&egrave;se qu&#39;evolution de PV ne soit pas monophyl&eacute;tique &agrave; travers tous les g&egrave;nes.

Verrues g&eacute;nitales chez les marsouins de Burmeister : caract&eacute;risation de Phocoena spinipinnis papillomavirus de type 1 (PsPV-1) et preuve pendant une seconde, vaguement apparent&eacute; PsPV.

Abbiamo individuato in sequenze da due papillomavirus lontanamente correlati in verruche genitali da focene spinipinne due, tra cui un esemplare di antigene-positivo PV e caratterizzato Phocoena spinipinnis papillomavirus type 1 (PsPV-1). Il genoma PsPV-1 comprende 7879 nt e presenta caratteristiche insolite. Manca un E7, un E8 e un bona fide E5 open reading frame (ORF) e ha un grande ORF E6. ORF L1 PsPV-1 ha mostrato la pi&ugrave; alta percentuale di identit&agrave; nucleotidica (54-55%) con tipo di papillomavirus umano 5, bovina papillomavirus tipo 3 (BPV-3) e Tursiops truncatus papillomavirus tipo 2 (TtPV-2). Questo garantisce che la classificazione di PsPV-1 come il prototipo del genere Omikronpapillomavirus. PsPV-1 cluster con TtPV-2 in E6 ed E1E2 alberi filogenetici e con-TtPV-2 e BPV-3 nell&#39;albero L2L1. Questo supporta l&#39;ipotesi che evoluzione PV non pu&ograve; essere monofiletico attraverso tutti i geni.

Le verruche genitali in focene spinipinne: caratterizzazione di Phocoena spinipinnis papillomavirus type 1 (PsPV-1) e le prove per un secondo, tiepidi PsPV.

Epidermodysplasia verruciformis (EV) is a genodermatosis associated with skin cancers that results from a selective susceptibility to related human papillomaviruses (EV HPV). Invalidating mutations in either of two genes (EVER1 and EVER2) with unknown functions cause most EV cases. We report that EVER1 and EVER2 proteins form a complex and interact with the zinc transporter 1 (ZnT-1), as shown by yeast two-hybrid screening, GST pull-down, and immunoprecipitation experiments. In keratinocytes, EVER and ZnT-1 proteins do not influence intracellular zinc concentration, but do affect intracellular zinc distribution. EVER2 was found to inhibit free zinc influx to nucleoli. Keratinocytes with a mutated EVER2 grew faster than wild-type keratinocytes. In transiently and stably transfected HaCaT cells, EVER and ZnT-1 down-regulated transcription factors stimulated by zinc (MTF-1) or cytokines (c-Jun and Elk), as detected with luciferase assays. To get some insight into the control of EV HPV infection, we searched for interaction between EVER and ZnT-1 and oncoproteins of cutaneous (HPV5) and genital (HPV16) genotypes. HPV16 E5 protein binds to EVER and ZnT-1 and blocks their negative regulation. The lack of a functional E5 protein encoded by EV HPV genome may account for host restriction of these viruses.

Regulation of cellular zinc balance as a potential mechanism of EVER-mediated protection against pathogenesis by cutaneous oncogenic human papillomaviruses.

&Eacute;pidermodysplasie verruciforme (EV) est une g&eacute;nodermatose associ&eacute;e aux cancers de la peau qui r&eacute;sulte d&#39;une sensibilit&eacute; s&eacute;lective &agrave; des virus du papillome humain (VPH EV). Mutations invalidantes dans un des deux g&egrave;nes (EVER1 et EVER2) avec fonctions inconnues causent la plupart des cas de EV. Les auteurs rapportent que les prot&eacute;ines EVER1 et EVER2 forment un complexe et interagissent avec le transporteur de zinc 1 (ZnT-1), tel qu&#39;illustr&eacute; par la levure-hybride d&eacute;pistage, TPS pull-down et des exp&eacute;riences d&#39;immunopr&eacute;cipitation. Dans les k&eacute;ratinocytes, les prot&eacute;ines EVER et ZnT-1 n&#39;influencent pas la concentration intracellulaire de zinc, mais affectent la distribution intracellulaire de zinc. EVER2 a &eacute;t&eacute; trouv&eacute; pour inhiber l&#39;afflux de zinc libre &agrave; nucl&eacute;oles. K&eacute;ratinocytes avec un EVER2 mut&eacute; a augment&eacute; plus rapidement que les k&eacute;ratinocytes de type sauvage. Dans transitoirement et stablement transfect&eacute; des cellules HaCaT, jamais et des facteurs de transcription diminu&eacute;es de ZnT-1 stimul&eacute;es par le zinc (MTF-1) ou de cytokines (c-Jun et le wapiti), comme d&eacute;tect&eacute;s avec des essais de la lucif&eacute;rase. Pour obtenir un aper&ccedil;u de la lutte contre l&#39;infection HPV EV, nous avons cherch&eacute; un interaction entre EVER et ZnT-1 et oncoprot&eacute;ines cutan&eacute;e (HPV5) et g&eacute;nitales (HPV 16) g&eacute;notypes. HPV16 E5 prot&eacute;ine se lie &agrave; EVER et ZnT-1 et bloque leur r&eacute;gulation n&eacute;gative. Restriction d&#39;h&ocirc;te de ces virus peut-&ecirc;tre expliquer l&#39;absence d&#39;une prot&eacute;ine fonctionnelle de E5 cod&eacute;e par le g&eacute;nome HPV EV.

R&egrave;glement du solde de zinc cellulaire comme un m&eacute;canisme potentiel de m&eacute;diation EVER protection contre pathogen&egrave;se par le virus du papillome humain oncog&egrave;ne cutan&eacute;.

Epidermodisplasia verruciforme (EV) &egrave; una genodermatosi associata a tumori della pelle che risulta da una suscettibilit&agrave; selettiva alle relative papillomavirus umano (HPV EV). Uno dei due geni (EVER1 ed EVER2) con funzioni sconosciute invalidanti le mutazioni causano la maggior parte dei casi di EV. Segnaliamo che EVER1 ed EVER2 proteine formano un complesso e interagiscano con il trasportatore di zinco 1 (ZnT-1), come mostrato da lievito due-ibrido screening, GST pull-down ed esperimenti di immunoprecipitazione. Nei cheratinociti, proteine sempre e ZnT-1 non influenza la concentrazione intracellulare di zinco, ma possono influenzare la distribuzione intracellulare di zinco. EVER2 &egrave; stato trovato per inibire lo zinco libero afflusso di nucleoli. Cheratinociti con un mutato EVER2 &egrave; cresciuto pi&ugrave; velocemente di cheratinociti wild-type. In transitoriamente e stabile transfected le cellule HaCaT, mai e fattori di trascrizione gi&ugrave;-regolato ZnT-1 stimolate da zinco (MTF-1) o citochine (c-Jun e alci), come rilevato con dosaggi di luciferasi. Per ottenere qualche intuizione nel controllo dell&#39;infezione da HPV EV, abbiamo cercato per interazione tra EVER e ZnT-1 e oncoproteins cutanea (HPV5) e genitale (HPV16) genotipi. HPV16 E5 proteina si lega a EVER e ZnT-1 e blocca la loro regolazione negativa. Restrizione di host di questi virus pu&ograve; spiegare la mancanza di una proteina E5 funzionale codificata dal genoma di HPV EV.

Regolazione dell&#39;equilibrio cellulare zinco come un potenziale meccanismo di protezione mediata da sempre contro la patogenesi di papillomavirus umano oncogenico cutaneo.

Infections by human papillomaviruses (HPVs) are the most frequently occurring sexually transmitted diseases. The crucial role of genital oncogenic HPV in cervical carcinoma development is now well established. In contrast, the role of cutaneous HPV in skin cancer development remains a matter of debate. Cutaneous beta-HPV strains show an amazing ubiquity. The fact that a few oncogenic genotypes cause cancers in patients suffering from epidermodysplasia verruciformis is in sharp contrast to the unapparent course of infection in the general population. Our recent investigations revealed that a natural barrier exists in humans, which protects them against infection with these papillomaviruses. A central role in the function of this HPV-specific barrier is played by a complex of the zinc-transporting proteins EVER1, EVER2, and ZnT-1, which maintain cellular zinc homeostasis. Apparently, the deregulation of the cellular zinc balance emerges as an important step in the life cycles not only of cutaneous but also of genital HPVs, although the latter viruses have developed a mechanism by which they can break the barrier and impose a zinc imbalance. Herein, we present a previously unpublished list of the cellular partners of EVER proteins, which points to future directions concerning investigations of the mechanisms of action of the EVER/ZnT-1 complex. We also present a general overview of the pathogenesis of HPV infections, taking into account the latest discoveries regarding the role of cellular zinc homeostasis in the HPV life cycle. We propose a potential model for the mechanism of function of the anti-HPV barrier.

The EVER proteins as a natural barrier against papillomaviruses: a new insight into the pathogenesis of human papillomavirus infections.

Les infections par le virus du papillome humain (HPV) sont les plus fr&Theta;quent maladies sexuellement transmissibles. Le r&ocirc;le crucial des VPH oncog&egrave;ne g&eacute;nital dans le d&eacute;veloppement du cancer du col est maintenant bien &eacute;tabli. En revanche, le r&ocirc;le des HPV cutan&eacute;e dans le d&eacute;veloppement du cancer de la peau reste un sujet de d&eacute;bat. B&ecirc;ta-HPV cutan&eacute;e souches pr&eacute;sentent une ubiquit&eacute; &eacute;tonnante. Le fait que quelques g&eacute;notypes oncog&egrave;nes causent des cancers chez les patients souffrant d&#39;epidermodysplasia verruciformis est en contraste avec le cours inapparente d&#39;infection dans la population g&eacute;n&eacute;rale. Nos investigations r&eacute;centes ont r&eacute;v&eacute;l&eacute; qu&#39;une barri&egrave;re naturelle existe chez les humains, qui les prot&egrave;ge contre l&#39;infection par ces virus du papillome. Un r&ocirc;le central dans le fonctionnement de cette barri&egrave;re de VPH sp&eacute;cifique est jou&eacute; par un complexe de prot&eacute;ines transport de zinc EVER1, EVER2 et ZnT-1, qui maintiennent l&#39;hom&eacute;ostasie cellulaire de zinc. Apparemment, la d&eacute;r&eacute;glementation de l&#39;&eacute;quilibre cellulaire zinc &eacute;merge comme une &eacute;tape importante dans le cycle de vie, non seulement de cutan&eacute;e, mais aussi du HPV g&eacute;nital, bien que les virus de ce dernier ont mis au point un m&eacute;canisme par lequel ils peuvent briser la barri&egrave;re et imposer un d&eacute;s&eacute;quilibre de zinc. Ici, nous pr&eacute;sentons une liste in&eacute;dite des partenaires cellulaires de jamais les prot&eacute;ines, qui pointe vers les orientations futures concernant les enqu&ecirc;tes sur les m&eacute;canismes d&#39;action du complexe EVER/ZnT-1. Nous pr&eacute;sentons &eacute;galement un aper&ccedil;u g&eacute;n&eacute;ral de la pathog&eacute;nie des infections &agrave; VPH, en tenant compte des derni&egrave;res d&eacute;couvertes concernant le r&ocirc;le de l&#39;hom&eacute;ostasie cellulaire de zinc dans le cycle de vie HPV. Nous proposons un mod&egrave;le pour le m&eacute;canisme de fonctionnement de la barri&egrave;re anti-VPH.

Les prot&eacute;ines EVER comme une barri&egrave;re naturelle contre le virus du papillome : un nouvel aper&ccedil;u de la pathogen&egrave;se des infections de virus du papillome humain.

Le infezioni da papillomavirus umano (HPV) sono le malattie sessualmente trasmesse pi&ugrave; frequentemente l&#39;avvenimento. Il ruolo cruciale dei genitali HPV oncogeni nello sviluppo del carcinoma cervicale &egrave; ora ben consolidato. Al contrario, il ruolo di HPV cutanee nello sviluppo del cancro della pelle rimane materia di dibattito. Ceppi beta-HPV cutanee mostrano un&#39;incredibile ubiquit&agrave;. Il fatto che alcuni genotipi oncogeni causano tumori in pazienti affetti da Epidermodisplasia verruciforme &egrave; in netto contrasto con il corso assenti di infezione nella popolazione generale. Nostre indagini recenti hanno rivelato che una barriera naturale esiste negli esseri umani, che li protegge contro l&#39;infezione con questi papillomavirus. Un ruolo centrale nella funzione di questa barriera specifici per HPV &egrave; interpretato da un complesso di proteine zinco-trasporto EVER1 EVER2 e ZnT-1, che mantenere l&#39;omeostasi cellulare di zinco. A quanto pare, la deregolamentazione dell&#39;equilibrio cellulare zinco emerge come un importante passo avanti nei cicli di vita non solo di cutaneo, ma anche di HPV genitale, anche se quest&#39;ultimo virus hanno sviluppato un meccanismo mediante il quale possono rompere la barriera e imporre uno squilibrio di zinco. Qui, vi presentiamo un elenco inedito dei partner del cellulare di mai le proteine, che punti a direzioni future riguardanti indagini dei meccanismi di azione del complesso EVER/ZnT-1. Inoltre presentiamo una panoramica generale della patogenesi di infezioni da HPV, prendendo in considerazione le ultime scoperte riguardo al ruolo dell&#39;omeostasi cellulare zinco nel ciclo di vita dell&#39;HPV. Vi proponiamo un potenziale modello per il meccanismo della funzione della barriera anti-HPV.

Le proteine sempre come una barriera naturale contro il papillomavirus: una nuova visione della patogenesi delle infezioni da papillomavirus umano.

Benchmarking a luciferase complementation assay for detecting protein complexes.

Analyse comparative d&#39;une compl&eacute;mentation de luciferase pour la d&eacute;tection des complexes prot&eacute;iques.

Analisi comparativa di un&#39;analisi di complementazione di luciferase per la rilevazione di complessi della proteina.

Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus that has been responsible for an epidemic outbreak of unprecedented magnitude in recent years. Since then, significant efforts have been made to better understand the biology of this virus, but we still have poor knowledge of CHIKV interactions with host cell components at the molecular level. Here we describe the extensive use of high-throughput yeast two-hybrid (HT-Y2H) assays to characterize interactions between CHIKV and human proteins. A total of 22 high-confidence interactions, which essentially involved the viral nonstructural protein nsP2, were identified and further validated in protein complementation assay (PCA). These results were integrated to a larger network obtained by extensive mining of the literature for reports on alphavirus-host interactions. To investigate the role of cellular proteins interacting with nsP2, gene silencing experiments were performed in cells infected by a recombinant CHIKV expressing Renilla luciferase as a reporter. Collected data showed that heterogeneous nuclear ribonucleoprotein K (hnRNP-K) and ubiquilin 4 (UBQLN4) participate in CHIKV replication in vitro. In addition, we showed that CHIKV nsP2 induces a cellular shutoff, as previously reported for other Old World alphaviruses, and determined that among binding partners identified by yeast two-hybrid methods, the tetratricopeptide repeat protein 7B (TTC7B) plays a significant role in this activity. Altogether, this report provides the first interaction map between CHIKV and human proteins and describes new host cell proteins involved in the replication cycle of this virus.

Mapping of Chikungunya virus interactions with host proteins identified nsP2 as a highly connected viral component.

Le virus Chikungunya (CHIKV) est un alphavirus transmises par la moustique qui a &eacute;t&eacute; responsable d&#39;une &eacute;pid&eacute;mie d&#39;une ampleur sans pr&eacute;c&eacute;dent ces derni&egrave;res ann&eacute;es. Depuis lors, des efforts importants ont &eacute;t&eacute; faits afin de mieux comprendre la biologie de ce virus, mais nous avons encore le manque de connaissances des interactions CHIKV avec des composants de cellule h&ocirc;te au niveau mol&eacute;culaire. Nous d&eacute;crivons ici la large utilisation du haut d&eacute;bit levure-hybride (HT-Y2H) essais pour caract&eacute;riser les interactions entre CHIKV et des prot&eacute;ines humaines. Un total de 22 haute-confiance des interactions, consistant essentiellement les prot&eacute;ines non structurales virale nsP2, ont &eacute;t&eacute; identifi&eacute;es et valid&eacute;s plus loin dans l&#39;analyse de compl&eacute;mentation prot&eacute;ique (PCA). Ces r&eacute;sultats ont &eacute;t&eacute; int&eacute;gr&eacute;s &agrave; un r&eacute;seau plus important obtenu par une vaste exploitation mini&egrave;re de la litt&eacute;rature pour les rapports sur les interactions h&ocirc;te-alphavirus. Afin d&#39;&eacute;tudier le r&ocirc;le des prot&eacute;ines cellulaires interagissant avec la nsP2, des exp&eacute;riences de g&egrave;ne silencieux ont &eacute;t&eacute; effectu&eacute;es dans les cellules infect&eacute;es par un CHIKV recombinant exprimant Renilla lucif&eacute;rase comme journaliste. A recueilli des donn&eacute;es a montr&eacute; que la ribonucl&eacute;oprot&eacute;ine nucl&eacute;aire h&eacute;t&eacute;rog&egrave;ne K (hnRNP-K) et ubiquilin 4 (UBQLN4) participent &agrave; la r&eacute;plication CHIKV in vitro. En outre, nous avons montr&eacute; que CHIKV nsP2 induit un cellulaire d&#39;arr&ecirc;t, comme d&eacute;j&agrave; signal&eacute; d&#39;autres alphavirus de l&#39;ancien monde et constate que parmi les partenaires de liaison identifi&eacute;s par des m&eacute;thodes de deux-hybride de levure, le tetratricopeptide r&eacute;p&egrave;te prot&eacute;ine 7 b (TTC7B) joue un r&ocirc;le important dans cette activit&eacute;. Au total, ce rapport fournit la premi&egrave;re carte de l&#39;interaction entre les CHIKV et les prot&eacute;ines humaines et d&eacute;crit les nouvelles prot&eacute;ines de cellules h&ocirc;tes impliqu&eacute;s dans le cycle de r&eacute;plication du virus.

Cartographie des interactions virus Chikungunya avec des prot&eacute;ines de l&#39;h&ocirc;te identifi&eacute; nsP2 comme une composante virale hautement connexe.

Chikungunya virus (CHIKV) &egrave; un alphavirus trasmesso da zanzare che &egrave; stato responsabile di un focolaio epidemico di ampiezza senza precedenti negli ultimi anni. Da allora, sono stati compiuti sforzi significativi per comprendere meglio la biologia di questo virus, ma abbiamo ancora scarsa conoscenza di interazioni CHIKV con componenti delle cellule ospite a livello molecolare. Qui descriviamo l&#39;uso estensivo di analisi di high-throughput lievito due-ibrido (HT-Y2H) per caratterizzare le interazioni tra CHIKV e proteine umane. Un totale di 22 interazioni di alta fiducia, che ha coinvolto essenzialmente il nsP2 virale proteine non strutturali, sono stati identificati e ulteriormente convalidato nell&#39;analisi della proteina di complementazione (PCA). Questi risultati sono stati integrati in una rete pi&ugrave; grande ottenuta dalla vasta mineraria della letteratura per i report sulle interazioni alphavirus-host. Per indagare il ruolo delle proteine cellulari interagendo con nsP2, sono stati eseguiti esperimenti del silenziamento del gene in cellule infettate da un CHIKV ricombinanti che esprimono Renilla luciferase come reporter. Dati raccolti hanno mostrato che ribonucleoproteina nucleare eterogenea K (hnRNP-K) e ubiquilin 4 (UBQLN4) partecipare alla replica di CHIKV in vitro. Inoltre, abbiamo dimostrato che CHIKV nsP2 induce un arresto cellulare, come precedentemente segnalato per altri alphaviruses del vecchio mondo e stabilito che tra i partner di associazione identificato da metodi di due-ibrido del lievito, del tetratricopeptide ripetere proteina 7B (TTC7B) svolge un ruolo significativo in questa attivit&agrave;. Complessivamente, questa relazione fornisce la prima mappa di interazione tra CHIKV e proteine umane e descrive il nuovo host cell proteine coinvolte nel ciclo di replicazione del virus.

Mappatura delle interazioni virus Chikungunya con proteine host identificato nsP2 come un componente virale altamente connessa.

Comparative interactomics is a strategy for inferring potential interactions among orthologous proteins or "interologs". Herein we focus, in contrast to standard homology-based inference, on the divergence of protein interaction profiles among closely related organisms, showing that the approach can correlate specific traits to phenotypic differences. As a model, this new comparative interactomic approach was applied at a large scale to human papillomaviruses (HPVs) proteins. The oncogenic potential of HPVs is mainly determined by the E6 and E7 early proteins. We have mapped and overlapped the virus-host protein interaction networks of E6 and E7 proteins from 11 distinct HPV genotypes, selected for their different tropisms and pathologies. We generated robust and comprehensive datasets by combining two orthogonal protein interaction assays: yeast two-hybrid (Y2H), and our recently described "high-throughput Gaussia princeps protein complementation assay" (HT-GPCA). HT-GPCA detects protein interaction by measuring the interaction-mediated reconstitution of activity of a split G. princeps luciferase. Hierarchical clustering of interaction profiles recapitulated HPV phylogeny and was used to correlate specific virus-host interaction profiles with pathological traits, reflecting the distinct carcinogenic potentials of different HPVs. This comparative interactomics constitutes a reliable and powerful strategy to decipher molecular relationships in virtually any combination of microorganism-host interactions.

Comparative analysis of virus-host interactomes with a mammalian high-throughput protein complementation assay based on Gaussia princeps luciferase.

Interactomique comparative est une strat&eacute;gie pour inf&eacute;rer les interactions potentielles entre les prot&eacute;ines orthologues ou &laquo; interologs &raquo;. Ici nous nous concentrons, contrairement &agrave; l&#39;inf&eacute;rence standard bas&eacute;e sur l&#39;homologie, sur la divergence des profils d&#39;interaction de prot&eacute;ines parmi les organismes apparent&eacute;s, montrant que l&#39;approche peut corr&eacute;ler des traits sp&eacute;cifiques aux diff&eacute;rences ph&eacute;notypiques. Comme un mod&egrave;le, cette nouvelle approche comparative interactomic a &eacute;t&eacute; appliqu&eacute;e &agrave; grande &eacute;chelle aux prot&eacute;ines de virus du papillome humain (HPV). Le potentiel oncog&egrave;ne du HPV est principalement d&eacute;termin&eacute; par les prot&eacute;ines pr&eacute;coces E6 et E7. Nous avons cartographi&eacute; et chevauchent les r&eacute;seaux d&#39;interactions prot&eacute;iques virus-h&ocirc;te de prot&eacute;ines E6 et E7 11 g&eacute;notypes de VPH distinctes, s&eacute;lectionn&eacute;s pour leurs diff&eacute;rents Tropismes et les pathologies. Nous avons g&eacute;n&eacute;r&eacute; des ensembles de donn&eacute;es robuste et complet en combinant deux essais d&#39;interaction de prot&eacute;ine orthogonale : levure-hybride (Y2H) et notre r&eacute;cemment d&eacute;crite &laquo; high-throughput Gaussia princeps prot&eacute;ine analyse de compl&eacute;mentation &raquo; (HT-ACPP). HT-ACPP d&eacute;tecte l&#39;interaction de la prot&eacute;ine en mesurant la reconstitution induite par l&#39;interaction de l&#39;activit&eacute; d&#39;un split g. princeps lucif&eacute;rase. Regroupement hi&eacute;rarchique des profils d&#39;interaction r&eacute;capitul&eacute; phylog&eacute;nie HPV et a &eacute;t&eacute; utilis&eacute; pour corr&eacute;ler les profils d&#39;interaction virus-h&ocirc;te sp&eacute;cifique avec des traits pathologiques, refl&eacute;tant les potentiels canc&eacute;rog&egrave;nes distinctes des diff&eacute;rents HPV. Ce comparatif interactomique constitue une strat&eacute;gie fiable et puissante pour d&eacute;chiffrer les relations mol&eacute;culaires dans pratiquement n&#39;importe quelle combinaison des interactions microorganismes-h&ocirc;te.

Analyse comparative d&#39;interactomes virus-h&ocirc;te avec un dosage de compl&eacute;mentation de mammif&egrave;res prot&eacute;ine haut d&eacute;bit bas&eacute;e sur Gaussia princeps lucif&eacute;rase.

Interattomica comparativa &egrave; una strategia per la deduzione di potenziali interazioni tra proteine orthologous o &quot;interologs&quot;. Qui ci concentriamo, in contrasto con standard inferenza basata su omologia, sulla divergenza dei profili di interazione della proteina tra organismi strettamente correlati, mostrando che l&#39;approccio pu&ograve; correlare specifici tratti di differenze fenotipiche. Come un modello, questo nuovo approccio comparativo interactomic &egrave; stato applicato a larga scala di proteine di papillomavirus umano (HPV). Il potenziale oncogenico del HPV &egrave; determinato principalmente dalle proteine E6 ed E7 precoce. Abbiamo mappato e le reti di interazione della proteina virus-ospite delle proteine E6 ed E7 da 11 diversi genotipi HPV, selezionati per la loro diverse tropismi e patologie si sovrappongono. Abbiamo generato robusto e completo set di dati dalla combinazione di due analisi di interazione proteina ortogonale: lievito due-ibrido (Y2H) e il nostro recentemente descritto &quot;high-throughput Gaussia princeps complementazione analisi della proteina&quot; (HT-GPCA). HT-GPCA rileva interazione proteina misurando la ricostituzione dell&#39;attivit&agrave; di un split g. princeps luciferase interazione mediata. Clustering gerarchico dei profili di interazione ricapitolate filogenesi HPV e fu utilizzato per correlare i profili di interazione virus-ospite specifico con tratti patologici, riflettendo i potenziali cancerogeni distinti di HPV diverso. Questo interattomica comparativa costituisce una strategia potente e affidabile per decifrare i rapporti molecolari in praticamente qualsiasi combinazione delle interazioni microrganismo-ospite.

Analisi comparativa dei virus-host interactomes con un dosaggio di complementazione dei mammiferi high-throughput di proteine basato su Gaussia princeps luciferasi.

Article	Year
Human papillomavirus type 5 E6 oncoprotein represses the transforming growth factor beta signaling pathway by binding to SMAD3. Journal of virology\| PubMed ID: 17020941	2006
Human papillomavirus in normal conjunctival tissue and in conjunctival papilloma: types and frequencies in a large series. The British journal of ophthalmology\| PubMed ID: 17166894	2007
Human papillomavirus: cause of epithelial lacrimal sac neoplasia? Acta ophthalmologica Scandinavica\| PubMed ID: 17362365	2007
Genital warts in Burmeister's porpoises: characterization of Phocoena spinipinnis papillomavirus type 1 (PsPV-1) and evidence for a second, distantly related PsPV. The Journal of general virology\| PubMed ID: 17554024	2007
Regulation of cellular zinc balance as a potential mechanism of EVER-mediated protection against pathogenesis by cutaneous oncogenic human papillomaviruses. The Journal of experimental medicine\| PubMed ID: 18158319	2008
The EVER proteins as a natural barrier against papillomaviruses: a new insight into the pathogenesis of human papillomavirus infections. Microbiology and molecular biology reviews : MMBR\| PubMed ID: 19487731	2009
Benchmarking a luciferase complementation assay for detecting protein complexes. Nature methods\| PubMed ID: 22127214	2011
Mapping of Chikungunya virus interactions with host proteins identified nsP2 as a highly connected viral component. Journal of virology\| PubMed ID: 22258240	2012
Comparative analysis of virus-host interactomes with a mammalian high-throughput protein complementation assay based on Gaussia princeps luciferase. Methods (San Diego, Calif.)\| PubMed ID: 22898364	2012

Patricia Cassonnet

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