title: Mapping of Chikungunya virus interactions with host proteins identified nsP2 as a highly connected viral component.
abstract: 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.
authors.affiliation: Unité de Génomique Virale et Vaccination, Institut Pasteur, CNRS URA 3015, Paris, France.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Bouraï
authors.forename: Marianne
authors.initials: M
authors.lastname: Lucas-Hourani
authors.forename: Hans Henrik
authors.initials: HH
authors.lastname: Gad
authors.forename: Christian
authors.initials: C
authors.lastname: Drosten
authors.forename: Yves
authors.initials: Y
authors.lastname: Jacob
authors.forename: Lionel
authors.initials: L
authors.lastname: Tafforeau
authors.forename: Patricia
authors.initials: P
authors.lastname: Cassonnet
authors.forename: Louis M
authors.initials: LM
authors.lastname: Jones
authors.forename: Delphine
authors.initials: D
authors.lastname: Judith
authors.forename: Thérèse
authors.initials: T
authors.lastname: Couderc
authors.forename: Marc
authors.initials: M
authors.lastname: Lecuit
authors.forename: Patrice
authors.initials: P
authors.lastname: André
authors.forename: Beate Mareike
authors.initials: BM
authors.lastname: Kümmerer
authors.forename: Vincent
authors.initials: V
authors.lastname: Lotteau
authors.forename: Philippe
authors.initials: P
authors.lastname: Desprès
authors.forename: Frédéric
authors.initials: F
authors.lastname: Tangy
authors.forename: Pierre-Olivier
authors.initials: PO
authors.lastname: Vidalain
mesh_terms: D018354:Alphavirus Infections / Q000378:metabolism* / Q000821:virology*; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D065632:Chikungunya Fever; D002646:Chikungunya virus / Q000235:genetics / Q000378:metabolism*; D034641:Heterogeneous-Nuclear Ribonucleoprotein K / Q000378:metabolism; D054884:Host-Pathogen Interactions; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism; D060066:Protein Interaction Maps; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D034641:Heterogeneous-Nuclear Ribonucleoprotein K; D009687:Nuclear Proteins; C421907:UBQLN4 protein, human; D017361:Viral Nonstructural Proteins