乙型肝炎病毒聚合酶序列所需的病毒RNA结合， RNA的包装，并

StephenW

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.

StephenW

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

摘要

乙型肝炎病毒通过使用多功能聚合酶（ HP）复制的DNA基因组通过的前基因组RNA（前基因组RNA ）逆转录。惠普的一个关键功能是它与病毒RNA信号特定关联，称为ε （ Hε ） ，位于前基因组RNA ，这是需要具体的前基因组RNA包装成病毒核衣壳和病毒反转录的起始。惠普利用自身作为蛋白引物（蛋白质吸）和Hε作为义不容辞的模板发起反转录。 HP是由四个结构域，包括末端蛋白（TP） ，间隔物，逆转录酶（RT），和RNA酶H结构域组成。最近发展， Hε依赖性，体外蛋白质吸测定用于本研究中，证明了几乎整个TP和RT域和大部分的RNA酶H结构域进行了所需蛋白灌注。内TP ，RT，和间隔的具体的残基被确定为适用于HP- Hε结合和/或蛋白质引发的关键。为Hε结合惠普顺序的要求相比，前基因组RNA的包装，和蛋白质吸允许惠普突变体的分类分为五个组，每组对这些复杂及相关流程截然不同的效果。对于惠普的这些相关的和必要的功能的HP要求的详细描述，将进一步阐明它的多种功能，并有助于这些功能的靶向抗病毒治疗的机制。
脚注

        收到2013年9月27日。
        接受二○一三年十一月一十日。
    地址对应剑鸣胡， [email protected] 。

    ↵ *现住址：曹峰，约翰·科克伦部，退伍军人事务医疗中心部，圣路易斯，密苏里州，美国。