title: Hepatitis C Virus Particle Assembly Involves Phosphorylation of NS5A by the c-Abl Tyrosine Kinase.
abstract: Hepatitis C virus (HCV) nonstructural protein 5A (NS5A) is thought to regulate the replication of viral RNA and the assembly of virus particles in a serine/threonine phosphorylation-dependent manner. However, the host kinases that phosphorylate NS5A have not been fully identified. Here, we show that HCV particle assembly involves the phosphorylation of NS5A by the c-Abl tyrosine kinase. Pharmacological inhibition or knockdown of c-Abl reduces the production of infectious HCV (J6/JFH1) particles in Huh-7.5 cells without markedly affecting viral RNA translation and replication. NS5A is tyrosine-phosphorylated in HCV-infected cells, and this phosphorylation is also reduced by the knockdown of c-Abl. Mutational analysis reveals that NS5A tyrosine phosphorylation is dependent, at least in part, on Tyr(330) (Tyr(2306) in polyprotein numbering). Mutation of this residue to phenylalanine reduces the production of infectious HCV particles but does not affect the replication of the JFH1 subgenomic replicon. These findings suggest that c-Abl promotes HCV particle assembly by phosphorylating NS5A at Tyr(330).
authors.affiliation: From the Division of Genome Science and Microbiology, Department of Pathological Sciences, Faculty of Medical Sciences, the Organization for Life Science Advancement Programs, University of Fukui, Fukui 910-1193, Japan and.
authors.forename: Shota
authors.initials: S
authors.lastname: Yamauchi
authors.affiliation: From the Division of Genome Science and Microbiology, Department of Pathological Sciences, Faculty of Medical Sciences, the Organization for Life Science Advancement Programs, University of Fukui, Fukui 910-1193, Japan and.
authors.forename: Kenji
authors.initials: K
authors.lastname: Takeuchi
authors.affiliation: From the Division of Genome Science and Microbiology, Department of Pathological Sciences, Faculty of Medical Sciences, the Organization for Life Science Advancement Programs, University of Fukui, Fukui 910-1193, Japan and.
authors.forename: Kazuyasu
authors.initials: K
authors.lastname: Chihara
authors.affiliation: From the Division of Genome Science and Microbiology, Department of Pathological Sciences, Faculty of Medical Sciences.
authors.forename: Xuedong
authors.initials: X
authors.lastname: Sun
authors.affiliation: From the Division of Genome Science and Microbiology, Department of Pathological Sciences, Faculty of Medical Sciences, the Third Department of Internal Medicine, Faculty of Medical Sciences, and.
authors.forename: Chisato
authors.initials: C
authors.lastname: Honjoh
authors.affiliation: From the Division of Genome Science and Microbiology, Department of Pathological Sciences, Faculty of Medical Sciences.
authors.forename: Hatsumi
authors.initials: H
authors.lastname: Yoshiki
authors.affiliation: the Division of Microbiology, Kobe University Graduate School of Medicine, Kobe 650-0017, Japan.
authors.forename: Hak
authors.initials: H
authors.lastname: Hotta
authors.affiliation: From the Division of Genome Science and Microbiology, Department of Pathological Sciences, Faculty of Medical Sciences, the Organization for Life Science Advancement Programs, University of Fukui, Fukui 910-1193, Japan and ksada@u-fukui.ac.jp.
authors.forename: Kiyonao
authors.initials: K
authors.lastname: Sada
mesh_terms: D000818:Animals; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D057809:HEK293 Cells; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D054884:Host-Pathogen Interactions; D006801:Humans; D015151:Immunoblotting; D018613:Microscopy, Confocal; D010766:Phosphorylation; D016315:Proto-Oncogene Proteins c-abl / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D019065:Virus Assembly; D012324:RNA-Dependent RNA Polymerase
chemical_list: D016315:Proto-Oncogene Proteins c-abl; D012367:RNA, Viral; D014443:Tyrosine; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase
keywords: ABL tyrosine kinase; hepatitis C virus (HCV); host-pathogen interaction; virology; virus assembly