人乙肝病毒聚合酶中HSP90结合位点的定位

Localization of HSP90 binding sites in the human hepatitis B virus polymerase.

作者信息

Cho G, Park S G, Jung G

机构信息

Department of Biology Education, Seoul National University, Seoul, 151-742, Korea.

出版信息

Biochem Biophys Res Commun. 2000 Mar 5;269(1):191-6. doi: 10.1006/bbrc.2000.2240.

DOI:10.1006/bbrc.2000.2240

PMID:10694498

Abstract

The fact that HSP90 proteins and their chaperonin partners play an important role in epsilon RNA binding of duck HBV Pol protein during duck HBV replication has been reported. To elucidate the molecular basis of HBV Pol/HSP90 interaction, we have characterized the HSP90 interaction to HBV Pol. We found that human HBV Pol protein upon synthesis in rabbit reticulocyte lysate formed a complex with HSP90 in vitro as duck HBV Pol did. In addition, HSP90 protein was copurified with MBP/POL protein expressed in HepG2 cells, suggesting that human HBV Pol protein is associated with HSP90 in vivo. To localize the HSP90 interaction site region, several deletion mutants of HBV Pol translated in vitro were immunoprecipitated with anti-HSP90 antibody. The result indicates that C-terminal regions of the TP and RT domains interact with HSP90 independently.

摘要

已有报道称，HSP90蛋白及其伴侣蛋白在鸭乙型肝炎病毒（duck HBV）复制过程中，对鸭HBV多聚酶（Pol）蛋白的εRNA结合起着重要作用。为阐明HBV Pol/HSP90相互作用的分子基础，我们对HSP90与HBV Pol的相互作用进行了表征。我们发现，在兔网织红细胞裂解物中合成的人HBV Pol蛋白，在体外与HSP90形成了复合物，就像鸭HBV Pol一样。此外，HSP90蛋白与在HepG2细胞中表达的MBP/POL蛋白共纯化，这表明人HBV Pol蛋白在体内与HSP90相关联。为定位HSP90相互作用位点区域，用抗HSP90抗体对体外翻译的几种HBV Pol缺失突变体进行了免疫沉淀。结果表明，TP和RT结构域的C末端区域独立与HSP90相互作用。