Mapping of Chikungunya virus interactions with host proteins identified nsP2 as a highly connected viral component
Autor(es): Bouraï Mehdi, Lucas-Hourani Marianne, Gad Hans Henrik, Drosten Christian, Jacob Yves, Tafforeau Lionel, Cassonnet Patricia, Jones Louis M, Judith Delphine, Couderc Thérèse, Lecuit Marc, André Patrice, Kümmerer Beate Mareike, Lotteau Vincent, Desprès Philippe, Tangy Frédéric, Vidalain Pierre-Olivier
Resumo: 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 underst- 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 - human proteins. A total of 22 high-confidence interactions, which essentially involved the viral nonstructural protein nsP2, were identified - 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) - 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, - 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 - human proteins - describes new host cell proteins involved in the replication cycle of this virus.
Imprenta: Journal of Virology, v. 86, n. 6, p. 3121-3134, 2012
Identificador do objeto digital: 10.1128/JVI.06390-11
Descritores: Chikungunya virus - Biosynthesis ; Chikungunya virus - Cell ; Chikungunya virus - Pathogenesis ; Chikungunya virus - Proteins ; Chikungunya virus - Viral infections ; Chikungunya virus - Molecular methods ; Chikungunya Virus - Virus ; Chikungunya virus - Chikungunya fever ; Chikungunya virus - Epidemic
Data de publicação: 2012
