Comparative Analysis of Hepatitis B Virus Polymerase Sequences Required for Viral RNA Binding, RNA Packaging, and Protein Priming
Scott A. Jones a,
Daniel N. Clark a,
Feng Cao b*,
John E. Tavis c and
Jianming Hu a
- Author Affiliations
a Department of Microbiology and Immunology, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania, USA
b VirRx, Inc., St. Louis, Missouri, USA
c Department of Molecular Microbiology and Immunology, Saint Louis University School of Medicine, Saint Louis, Missouri, USA
ABSTRACT
Hepatitis B virus replicates a DNA genome through reverse transcription of a pregenomic RNA (pgRNA) by using a multifunctional polymerase (HP). A critical function of HP is its specific association with a viral RNA signal, termed ε (Hε), located on pgRNA, which is required for specific packaging of pgRNA into viral nucleocapsids and initiation of viral reverse transcription. HP initiates reverse transcription by using itself as a protein primer (protein priming) and Hε as the obligatory template. HP is made up of four domains, including the terminal protein (TP), the spacer, the reverse transcriptase (RT), and the RNase H domains. A recently developed, Hε-dependent, in vitro protein priming assay was used in this study to demonstrate that almost the entire TP and RT domains and most of the RNase H domain were required for protein priming. Specific residues within TP, RT, and the spacer were identified as being critical for HP-Hε binding and/or protein priming. Comparison of HP sequence requirements for Hε binding, pgRNA packaging, and protein priming allowed the classification of the HP mutants into five groups, each with distinct effects on these complex and related processes. Detailed characterization of HP requirements for these related and essential functions of HP will further elucidate the mechanisms of its multiple functions and aid in the targeting of these functions for antiviral therapy.
FOOTNOTES
Received 27 September 2013.
Accepted 10 November 2013.
Address correspondence to Jianming Hu, [email protected].
↵* Present address: Feng Cao, John Cochran Division, Department of Veteran's Affairs Medical Center, St. Louis, Missouri, USA.
作者: StephenW 时间: 2014-1-15 11:22
乙型肝炎病毒聚合酶序列所需的病毒RNA结合, RNA的包装,并引发蛋白质的比较分析
斯科特A.琼斯,
丹尼尔· N.克拉克,
曹峰B * ,
约翰E的Tavis c和
建明胡一
- 作者所属机构
微生物学与免疫学,医学,好时,宾夕法尼亚州,美国宾夕法尼亚州立大学的一个系
b VirRx公司,圣路易斯,密苏里州,美国
分子微生物学与免疫学,圣路易斯大学医学院,圣路易斯,密苏里州,美国的Ç部