Two defective forms of reverse transcriptase can complement to restore retroviral infectivity.
Two defective forms of reverse transcriptase can complement to restore retroviral infectivity.
复制标题
两种有缺陷的逆转录酶可以互补以恢复逆转录病毒的感染性。
DOI：
10.1002/j.1460-2075.1993.tb06128.x
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Goff,SP
中科院分区：
文献类型：
--
作者：
Telesnitsky,A;Goff,SP
逆转录病毒DNA合成需要DNA聚合酶和逆转录酶（RT）的RNaseH活性。为了测试两个有缺陷的RT--一个在RNaseH结构域中携带突变，另一个在DNA聚合酶中携带突变--是否可以一起工作以完成病毒DNA合成，我们产生了含有两种突变RT的莫洛尼鼠白血病病毒（M-MuLV）的表型混合病毒体。一个RNaseH催化位点突变体补充了两个测试的DNA聚合酶突变体和少量的完整的病毒DNA产生。这表明，当DNA聚合酶和RNaseH活性由单独的RT分子提供时，逆转录病毒DNA合成可以完成-尽管效率不高。其他RNaseH突变体未能互补，这表明RNaseH结构域的某些方面对RT的DNA聚合酶功能至关重要。表型混合的病毒体也被用来证明RT和整合酶（IN）可以提供单独的多蛋白前体和完成逆转录病毒复制的早期阶段。
Retroviral DNA synthesis requires both the DNA polymerase and the RNaseH activities of reverse transcriptase (RT). To test whether two defective RTs‐‐one carrying a mutation in the RNaseH domain and the other with a mutation in DNA polymerase‐‐could work together to complete viral DNA synthesis, we generated phenotypically mixed virions of Moloney murine leukemia virus (M‐MuLV) that contained two kinds of mutant RTs. One RNaseH catalytic site mutant complemented both tested DNA polymerase mutants and small amounts of intact viral DNA were generated. This demonstrates that retroviral DNA synthesis can be completed‐‐albeit inefficiently‐‐when DNA polymerase and RNaseH activities are provided by separate RT molecules. Other RNaseH mutants failed to complement, suggesting that some aspects of the RNaseH domain are essential to RT's DNA polymerase function. Phenotypically mixed virions were also used to demonstrate that RT and integrase (IN) can be provided by separate polyprotein precursors and complete the early stages of retroviral replication.