Domain structure of the Moloney murine leukemia virus reverse transcriptase: mutational analysis and separate expression of the DNA polymerase and RNase H activities.
Domain structure of the Moloney murine leukemia virus reverse transcriptase: mutational analysis and separate expression of the DNA polymerase and RNase H activities.
复制标题
莫洛尼鼠白血病病毒逆转录酶的结构域：突变分析以及 DNA 聚合酶和 RNase H 活性的单独表达。
DOI：
10.1073/pnas.85.6.1777
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发表时间：
1988
影响因子：
11.1
通讯作者：
Goff,SP
中科院分区：
文献类型：
--
作者：
Tanese,N;Goff,SP
莫洛尼鼠白血病病毒的逆转录酶与所有逆转录病毒的逆转录酶一样，表现出能够在RNA或DNA模板上合成的DNA聚合酶活性和对RNA.DNA杂合体形式的RNA具有特异性的RNA酶H活性。我们已经产生了细菌中表达的莫洛尼鼠白血病病毒酶的接头插入突变体的文库，并测定了这些突变体的两种酶活性。影响DNA聚合酶活性的突变集中在基因的5 '端近三分之二处，而影响RNase H的突变集中在剩余的3'端近三分之一处。基于这些图谱，制备了分别表达每个结构域的质粒;对这些质粒编码的蛋白质的测定表明，每个结构域仅表现出预期的活性。
The reverse transcriptase of Moloney murine leukemia virus, like that of all retroviruses, exhibits a DNA polymerase activity capable of synthesis on RNA or DNA templates and an RNase H activity with specificity for RNA in the form of an RNA.DNA hybrid. We have generated a library of linker insertion mutants of the Moloney murine leukemia virus enzyme expressed in bacteria and assayed these mutants for both enzymatic activities. Those mutations affecting the DNA polymerase activity were clustered in the 5'-proximal two-thirds of the gene, and those affecting RNase H were in the remaining 3' one-third. Based on these maps, plasmids were made that expressed each one of the domains separately; assays of the proteins encoded by these plasmids showed that each domain exhibited only the expected activity.