title: PML tumor suppressor protein is required for HCV production.
abstract: PML tumor suppressor protein, which forms discrete nuclear structures termed PML-nuclear bodies, has been associated with several cellular functions, including cell proliferation, apoptosis and antiviral defense. Recently, it was reported that the HCV core protein colocalizes with PML in PML-NBs and abrogates the PML function through interaction with PML. However, role(s) of PML in HCV life cycle is unknown. To test whether or not PML affects HCV life cycle, we examined the level of secreted HCV core and the infectivity of HCV in the culture supernatants as well as the level of HCV RNA in HuH-7-derived RSc cells, in which HCV-JFH1 can infect and efficiently replicate, stably expressing short hairpin RNA targeted to PML. In this context, the level of secreted HCV core and the infectivity in the supernatants from PML knockdown cells was remarkably reduced, whereas the level of HCV RNA in the PML knockdown cells was not significantly affected in spite of very effective knockdown of PML. In fact, we showed that PML is unrelated to HCV RNA replication using the subgenomic HCV-JFH1 replicon RNA, JRN/3-5B. Furthermore, the infectivity of HCV-like particle in the culture supernatants was significantly reduced in PML knockdown JRN/3-5B cells expressing core to NS2 coding region of HCV-JFH1 genome using the trans-packaging system. Finally, we also demonstrated that INI1 and DDX5, the PML-related proteins, are involved in HCV production. Taken together, these findings suggest that PML is required for HCV production.
authors.affiliation: Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, 2-5-1, Shikata-cho, Okayama 700-8558, Japan.
authors.forename: Misao
authors.initials: M
authors.lastname: Kuroki
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Ariumi
authors.forename: Makoto
authors.initials: M
authors.lastname: Hijikata
authors.forename: Masanori
authors.initials: M
authors.lastname: Ikeda
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Dansako
authors.forename: Takaji
authors.initials: T
authors.lastname: Wakita
authors.forename: Kunitada
authors.initials: K
authors.lastname: Shimotohno
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Kato
mesh_terms: D002460:Cell Line; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000502:physiology; D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000502:physiology; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology; D055785:Gene Knockdown Techniques; D016174:Hepacivirus / Q000502:physiology*; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics / Q000502:physiology*; D000071156:Promyelocytic Leukemia Protein; D012367:RNA, Viral / Q000096:biosynthesis*; D000071796:SMARCB1 Protein; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000502:physiology*; D014779:Virus Replication
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D012367:RNA, Viral; D000071796:SMARCB1 Protein; C513266:SMARCB1 protein, human; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C070576:PML protein, human; C506749:Ddx5 protein, human; D053487:DEAD-box RNA Helicases