Direct interaction between alpha-actinin and hepatitis C virus NS5B.
It has been suggested that cellular proteins are involved in hepatitis C virus (HCV) RNA replication. By using the yeast two-hybrid system, we isolated seven cDNA clones encoding proteins interacting with HCV RNA polymerase (NS5B) from a human liver cDNA library. For one of these, alpha-actinin, we confirmed the interaction by coimmunoprecipitation, immunofluorescent staining and confocal microscopic analysis. Experiments with deletion mutants showed that domains NS5B(84-95), NS5B(466-478), and alpha-actinin(621-733) are responsible for the interaction. Studies of the HCV subgenomic replicon system with small interference RNA indicate that alpha-actinin is essential for HCV RNA replication. Our results suggest alpha-actinin may be a component of the HCV replication complex.
Lan
Shuiyun
S
Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, Shanghai 200032, PR China.
Wang
Hua
H
Jiang
Hong
H
Mao
Hongxia
H
Liu
Xiaoying
X
Zhang
Xiaonan
X
Hu
Yunwen
Y
Xiang
Li
L
Yuan
Zhenghong
Z
D000185:Actinin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D015723:Gene Library; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D018613:Microscopy, Confocal; D011233:Precipitin Tests; D017434:Protein Structure, Tertiary; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D012093:Replicon / Q000235:genetics; D017384:Sequence Deletion; D020798:Two-Hybrid System Techniques; D014779:Virus Replication / Q000187:drug effects / Q000502:physiology
D034741:RNA, Small Interfering; D012367:RNA, Viral; D000185:Actinin; D012321:DNA-Directed RNA Polymerases