title: Interaction of hepatitis B virus X protein with PARP1 results in inhibition of DNA repair in hepatocellular carcinoma.
abstract: Hepatitis B virus X protein (HBx) contributes to the development of hepatocellular carcinoma (HCC), probably by regulating activities of many host or viral proteins through protein-protein interactions. In this study, we identified poly(ADP-ribose) polymerase (PARP1), a crucial factor in DNA repair, as an HBx-interacting protein using a proteomics approach. Coimmunoprecipitation and proximity ligation assays confirmed the binding and colocalization of HBx and PARP1 in the nucleus. The carboxyl-terminus of HBx protein bound to the catalytic domain of PARP1, and this binding reduced the enzymatic activity of PARP1 in both in vitro and in vivo assays. HBx interrupted the binding of PARP1 to Sirt6, which catalyzes the mono-ADP-ribosylation required for DNA repair. Consistently, overexpression of HBx inhibited the clearance of γH2AX DNA repair foci generated under oxidative stress in Chang liver cells. Recruitment of the DNA repair complex to the site-specific double-strand breaks was inhibited in the presence of HBx, when measured by laser microirradiation assay and damage-specific chromatin immunoprecipitation assays. Consequently, HBx increased signs of DNA damage such as accumulation of 8-hydroxy-2'-deoxyguanosine and comet formation, which were reversed by overexpression of PARP1 and/or Sirt6. Finally, the interaction between PARP1 and Sirt6 was markedly lower in the livers of HBx-transgenic mice and specimens obtained from HCC patients to compare with the corresponding control. Our data suggest that the physical interaction of HBx and PARP1 accelerates DNA damage by inhibiting recruitment of the DNA repair complex to the damaged DNA sites, which may lead to the onset of hepatocarcinogenesis.
authors.affiliation: College of Pharmacy and Bio-MAX Institute, Seoul National University, Seoul, Korea.
authors.forename: T-Y
authors.initials: TY
authors.lastname: Na
authors.affiliation: College of Pharmacy and Bio-MAX Institute, Seoul National University, Seoul, Korea.
authors.forename: N-L
authors.initials: NL
authors.lastname: Ka
authors.affiliation: Department of Pathology, Integrated Genomic Research Center for Metabolic Regulation, Yonsei University College of Medicine, Seoul, Korea.
authors.forename: H
authors.initials: H
authors.lastname: Rhee
authors.affiliation: College of Veterinary Medicine, Interdisciplinary Program for Bioinformatics and Program for Cancer Biology, Seoul National University and Korea Mouse Phenotyping Center, Seoul, Korea.
authors.forename: D
authors.initials: D
authors.lastname: Kyeong
authors.affiliation: College of Pharmacy and Bio-MAX Institute, Seoul National University, Seoul, Korea.
authors.forename: M-H
authors.initials: MH
authors.lastname: Kim
authors.affiliation: College of Veterinary Medicine, Interdisciplinary Program for Bioinformatics and Program for Cancer Biology, Seoul National University and Korea Mouse Phenotyping Center, Seoul, Korea.
authors.forename: J K
authors.initials: JK
authors.lastname: Seong
authors.affiliation: Department of Pathology, Integrated Genomic Research Center for Metabolic Regulation, Yonsei University College of Medicine, Seoul, Korea.|Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Korea.
authors.forename: Y N
authors.initials: YN
authors.lastname: Park
authors.affiliation: College of Pharmacy and Bio-MAX Institute, Seoul National University, Seoul, Korea.
authors.forename: M-O
authors.initials: MO
authors.lastname: Lee
mesh_terms: D000818:Animals; D063646:Carcinogenesis / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000473:pathology / Q000821:virology; D004249:DNA Damage / Q000235:genetics; D004260:DNA Repair / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000235:genetics / Q000472:pathogenicity; D006657:Histones / Q000235:genetics; D006801:Humans; D008113:Liver Neoplasms / Q000000981:diagnostic imaging / Q000235:genetics* / Q000473:pathology / Q000821:virology; D051379:Mice; D008822:Mice, Transgenic; D000071137:Poly (ADP-Ribose) Polymerase-1 / Q000235:genetics* / Q000378:metabolism; D037761:Sirtuins / Q000235:genetics* / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D054334:Viral Regulatory and Accessory Proteins
chemical_list: D004268:DNA-Binding Proteins; C468783:H2AX protein, human; D006657:Histones; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein; C499320:PARP1 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; C516592:SIRT6 protein, human; D037761:Sirtuins