Subproteomic study of hepatitis C virus replicon reveals Ras-GTPase-activating protein binding protein 1 as potential HCV RC component.
Hepatitis C virus (HCV) RNA synthesis takes place on a detergent resistant membrane (DRM) structure. To identify potential cellular proteins related to HCV replication complexes (RC), we purified DRMs from HCV subgenomic replicon cells and its parental Huh7 cells. The proteins of DRM fractions were separated by two-dimensional gel electrophoresis and identified by mass spectrometry. Comparing with parental Huh7 cells, 60 proteins were up-regulated while 14 proteins were down-regulated in HCV replicon cells. Ras-GTPase-activating protein binding protein 1 (G3BP1), one of the elevated proteins, was found to be associated with HCV NS5B and knockdown of G3BP1 by siRNA in HCV replicon cells significantly reduced HCV replication, which may indicate it a potential component of HCV RC. These results suggest that HCV viral gene and proteins may regulate the presence of host cellular proteins in DRM, ensure appropriate concentrations of replication components, and hence control the rates or efficiencies of HCV replication.
Yi
Zhigang
Z
Key Laboratory of Medical Molecular Virology, Institute of Medical Microbiology, Shanghai Medical College, Fudan University, Shanghai 200032, China.
Fang
Caiyun
C
Pan
Tingting
T
Wang
Jiadong
J
Yang
Pengyuan
P
Yuan
Zhenghong
Z
D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects; D004265:DNA Helicases; D003902:Detergents / Q000494:pharmacology; D016174:Hepacivirus / Q000187:drug effects / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D000075223:Poly-ADP-Ribose Binding Proteins; D040901:Proteomics; D020365:RNA Helicases; D000067776:RNA Recognition Motif Proteins; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014779:Virus Replication
D002352:Carrier Proteins; D003902:Detergents; D000075223:Poly-ADP-Ribose Binding Proteins; D000067776:RNA Recognition Motif Proteins; D004265:DNA Helicases; C116070:G3BP1 protein, human; D020365:RNA Helicases