Engelman A, Bushman F D, Craigie R
Laboratory of Molecular Biology, National Institute of Diabetes, Digestive and Kidney Diseases, Bethesda, MD 20892.
EMBO J. 1993 Aug;12(8):3269-75. doi: 10.1002/j.1460-2075.1993.tb05996.x.
HIV-1 integrase protein possesses the 3' processing and DNA strand transfer activities that are required to integrate HIV DNA into a host chromosome. The N-, C-terminal and core domains of integrase are necessary for both activities in vitro. We find that certain pairs of mutant integrase proteins, which are inactive when each protein is assayed alone, can support near wild type levels of activity when both proteins are present together in the reaction mixture. This complementation implies that HIV-1 integrase functions as a multimer and has enabled us to probe the organization of the functional domains within active mixed multimers. We have identified a minimal set of functional integrase domains that are sufficient for 3' processing and DNA strand transfer and find that some domains are contributed in trans by separate monomers within the functional complex.
HIV-1整合酶蛋白具有将HIV DNA整合到宿主染色体中所需的3'加工和DNA链转移活性。整合酶的N端、C端和核心结构域在体外对这两种活性都是必需的。我们发现，某些成对的突变整合酶蛋白，当单独检测每种蛋白时是无活性的，但当两种蛋白同时存在于反应混合物中时，它们能够支持接近野生型水平的活性。这种互补作用表明HIV-1整合酶作为一种多聚体发挥作用，并且使我们能够探究活性混合多聚体内功能结构域的组织方式。我们已经鉴定出一组最小的功能性整合酶结构域，它们足以进行3'加工和DNA链转移，并且发现一些结构域是由功能复合物中的不同单体反式提供的。
