Chaperonin TRiC/CCT participates in replication of hepatitis C virus genome via interaction with the viral NS5B protein

• Published in: Virology (New York, N.Y.) Vol. 410; no. 1; pp. 38 - 47
• Main Authors: Inoue, Yasushi, Aizaki, Hideki, Hara, Hiromichi, Matsuda, Mami, Ando, Tomomi, Shimoji, Tetsu, Murakami, Kyoko, Masaki, Takahiro, Shoji, Ikuo, Homma, Sakae, Matsuura, Yoshiharu, Miyamura, Tatsuo, Wakita, Takaji, Suzuki, Tetsuro
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 05.02.2011
• Subjects: antibodies; Cell Line, Tumor; Chaperonin; Chaperonins - metabolism; DNA-directed RNA polymerase; Gene Expression Regulation, Viral - physiology; genome; Hepacivirus - genetics; Hepacivirus - physiology; Hepatitis C virus; Hepatocytes - virology; Humans; Infectious Disease; Non-structural protein; polypeptides; proteome; Replication; RNA; RNA, Viral - genetics; RNA, Viral - metabolism; Viral Core Proteins - genetics; Viral Core Proteins - metabolism; viral nonstructural proteins; Viral Nonstructural Proteins - genetics; Viral Nonstructural Proteins - metabolism; Virus Replication - physiology; viruses; Chaperonin; Replication; Hepatitis C virus; Non-structural protein
• ISSN: 0042-6822; 1096-0341; 1096-0341
• DOI: 10.1016/j.virol.2010.10.026

To identify the host factors implicated in the regulation of hepatitis C virus (HCV) genome replication, we performed comparative proteome analyses of HCV replication complex (RC)-rich membrane fractions prepared from cells harboring genome-length bicistronic HCV RNA at the exponential and stationary growth phases. We found that the eukaryotic chaperonin T-complex polypeptide 1 (TCP1)-ring complex/chaperonin-containing TCP1 (TRiC/CCT) plays a role in the replication possibly through an interaction between subunit CCT5 and the viral RNA polymerase NS5B. siRNA-mediated knockdown of CCT5 suppressed RNA replication and production of the infectious virus. Gain-of-function activity was shown following co-transfection with whole eight TRiC/CCT subunits. HCV RNA synthesis was inhibited by an anti-CCT5 antibody in a cell-free assay. These suggest that recruitment of the chaperonin by the viral nonstructural proteins to the RC, which potentially facilitate folding of the RC component(s) into the mature active form, may be important for efficient replication of the HCV genome.