RECONSTITUTION INVITRO OF RNASE-H ACTIVITY BY USING PURIFIED N-TERMINAL AND C-TERMINAL DOMAINS OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 REVERSE-TRANSCRIPTASE
RECONSTITUTION INVITRO OF RNASE-H ACTIVITY BY USING PURIFIED N-TERMINAL AND C-TERMINAL DOMAINS OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 REVERSE-TRANSCRIPTASE
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DOI：
10.1073/pnas.88.4.1148
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发表时间：
1991-02-01
影响因子：
11.1
通讯作者：
NODES, BR
中科院分区：
文献类型：
--
作者：
HOSTOMSKY, Z;HOSTOMSKA, Z;NODES, BR
分别表达并纯化了人类免疫缺陷病毒1型逆转录酶的两个组成蛋白结构域。N-末端结构域(P51)是一种对盐敏感的DNA聚合酶活性的单体蛋白。C末端结构域(P15)本身没有可检测到的RNaseH活性。然而，分离的p51和p15在体外的结合导致了RNaseH活性在特定底物上的重建。这些结果表明，HIV-1逆转录酶的结构域在功能上的相互依赖性比Moloney鼠白血病病毒的逆转录酶要高得多。
Two constituent protein domains of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase were expressed separately and purified to homogeneity. The N-terminal domain (p51) behaves as a monomeric protein exhibiting salt-sensitive DNA polymerase activity. The C-terminal domain (p15) on its own has no detectable RNase H activity. However, the combination of both isolated p51 and p15 in vitro leads to reconstitution of RNase H activity on a defined substrate. These results demonstrate that domains of HIV-1 reverse transcriptase are functionally interdependent to a much higher degree than in the case of reverse transcriptase from Moloney murine leukemia virus.