Prasad V R, Goff S P
Department of Biochemistry, Columbia University, College of Physicians and Surgeons, New York, NY 10032.
Proc Natl Acad Sci U S A. 1989 May;86(9):3104-8. doi: 10.1073/pnas.86.9.3104.
A plasmid construct expressing the p66 version of the human immunodeficiency virus reverse transcriptase as a bacterial fusion protein was subjected to in vitro mutagenesis, and the resulting variant proteins were assayed to define the locations of the two major enzymatic activities. The DNA polymerase activity was localized to the N-terminal portion of the protein; mutations altering or eliminating the C-terminal portion had little or no effect on that activity. The results suggest that, in contrast with previous reports, the p51 subunit found in virions should exhibit DNA polymerase activity. Mutations in many parts of the protein eliminated RNase H activity, suggesting that several areas are needed for proper folding and generation of that activity.
一个表达人免疫缺陷病毒逆转录酶p66版本作为细菌融合蛋白的质粒构建体接受了体外诱变，对产生的变体蛋白进行了测定以确定两种主要酶活性的位置。DNA聚合酶活性定位于该蛋白的N端部分；改变或消除C端部分的突变对该活性几乎没有影响。结果表明，与之前的报道相反，在病毒颗粒中发现的p51亚基应具有DNA聚合酶活性。该蛋白许多部位的突变消除了核糖核酸酶H活性，这表明需要几个区域来实现该活性的正确折叠和产生。
