MUTATIONS IN THE EPSILON-SEQUENCES OF HUMAN HEPATITIS-B VIRUS AFFECT BOTH RNA ENCAPSIDATION AND REVERSE TRANSCRIPTION
MUTATIONS IN THE EPSILON-SEQUENCES OF HUMAN HEPATITIS-B VIRUS AFFECT BOTH RNA ENCAPSIDATION AND REVERSE TRANSCRIPTION
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DOI：
10.1128/jvi.69.5.3067-3073.1995
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发表时间：
1995-05-01
影响因子：
5.4
通讯作者：
GOFF, SP
中科院分区：
文献类型：
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作者：
FALLOWS, DA;GOFF, SP
嗜肝DNA病毒通过感染肝细胞的细胞质中亚病毒核心颗粒内的RNA中间体的逆转录进行复制。病毒聚合酶对位于前基因组RNA的5'端的腺苷酸化信号的识别发生在核心颗粒组装的早期。该序列包含一组嵌套的反向重复序列，其形成稳定的茎环结构，显示在RNA包装中发挥作用，并且最近被认为是负链DNA合成的起始位点。我们已经引入了各种定点突变到人类B型肝炎病毒的DNA序列，以研究它们对转染HuH7细胞中病毒复制的影响。我们已经确定了两类突变：对包装产生不利影响的突变和包装RNA但对DNA合成产生不利影响的突变。对这些突变体的分析使我们能够确定在RNA包装和逆转录过程中起作用的顺式作用信号的单独特征。
Hepadnaviruses replicate by reverse transcription of an RNA intermediate within subviral core particles in the cytoplasm of infected hepatocytes. Recognition of the epsilon encapsidation signal located on the 5' end of the pregenomic RNA by the viral polymerase occurs early in core particle assembly. The epsilon sequences contain a set of nested inverted repeats which form a stable stem-loop structure shown to play a role in RNA packaging and recently implicated as the site of initiation of minus-strand DNA synthesis. We have introduced a variety of site-directed mutations into the epsilon sequences of human hepatitis B virus to study their effects on viral replication in transfected HuH7 cells. We have identified two classes of mutations: those which adversely affect packaging and those which package RNA but adversely affect DNA synthesis. Analysis of these mutants has allowed us to identify separate features of the epsilon cis-acting signal which function in the processes of RNA packaging and reverse transcription.