Involvement of creatine kinase B in hepatitis C virus genome replication through interaction with the viral NS4A protein.
Persistent infection with hepatitis C virus (HCV) is a major cause of chronic liver diseases. The aim of this study was to identify host cell factor(s) participating in the HCV replication complex (RC) and to clarify the regulatory mechanisms of viral genome replication dependent on the host-derived factor(s) identified. By comparative proteome analysis of RC-rich membrane fractions and subsequent gene silencing mediated by RNA interference, we identified several candidates for RC components involved in HCV replication. We found that one of these candidates, creatine kinase B (CKB), a key ATP-generating enzyme that regulates ATP in subcellular compartments of nonmuscle cells, is important for efficient replication of the HCV genome and propagation of infectious virus. CKB interacts with HCV NS4A protein and forms a complex with NS3-4A, which possesses multiple enzyme activities. CKB upregulates both NS3-4A-mediated unwinding of RNA and DNA in vitro and replicase activity in permeabilized HCV replicating cells. Our results support a model in which recruitment of CKB to the HCV RC compartment, which has high and fluctuating energy demands, through its interaction with NS4A is important for efficient replication of the viral genome. The CKB-NS4A association is a potential target for the development of a new type of antiviral therapeutic strategy.
Hara
Hiromichi
H
Department of Virology II, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
Aizaki
Hideki
H
Matsuda
Mami
M
Shinkai-Ouchi
Fumiko
F
Inoue
Yasushi
Y
Murakami
Kyoko
K
Shoji
Ikuo
I
Kawakami
Hayato
H
Matsuura
Yoshiharu
Y
Lai
Michael M C
MM
Miyamura
Tatsuo
T
Wakita
Takaji
T
Suzuki
Tetsuro
T
D000255:Adenosine Triphosphate / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D052276:Creatine Kinase, BB Form / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral; D016679:Genome, Viral; D016174:Hepacivirus / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D006526:Hepatitis C / Q000378:metabolism / Q000821:virology; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D040901:Proteomics; D034622:RNA Interference; D012367:RNA, Viral / Q000235:genetics; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; C455041:NS4A cofactor peptide, Hepatitis C virus; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; D000255:Adenosine Triphosphate; D052276:Creatine Kinase, BB Form