Crystal structure of the HIV-1 integrase catalytic core and C-terminal domains: A model for viral DNA binding
Crystal structure of the HIV-1 integrase catalytic core and C-terminal domains: A model for viral DNA binding
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DOI：
10.1073/pnas.150220297
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发表时间：
2000-07-18
影响因子：
11.1
通讯作者：
Stroud, RM
中科院分区：
文献类型：
--
作者：
Chen, JCH;Krucinski, J;Stroud, RM
全长HIV-1整合酶(IN)的不溶性限制了以前对单个结构域的结构分析，通过引入五个点突变，我们设计了一种更可溶的IN，使我们能够产生多结构域的HIV-1 IN晶体。报道了第一个多结构域HIV-1 IN结构。它包含催化核心区和C-末端结构域(残基52-288)。分解到2.8埃的结构是一个Y形二聚体，在二聚体中，催化核心区形成唯一的二聚体界面，C-末端结构域相距55埃，26-AAα-螺旋，α6，连接C-末端结构域和催化核心。两个α6螺旋中的一个在一个已知的蛋白水解点附近发生扭结，表明它可能在整合过程中作为一个灵活的肘部重新定位结构域。两个以序列无关的方式与DNA结合的蛋白质在结构上与HIV-1的IN C-末端结构域同源，这表明蛋白质与DNA的相互作用类似，在整合过程中，IN C-末端结构域可能起到结合、弯曲和定位病毒DNA的作用，两个单体贡献的一条带正电荷的氨基酸出现在二聚体的每个活性部位，表明每个病毒DNA末端都有一个最小的二聚体平台。分离的催化核心域(残基52-210)的晶体结构在1.6埃分辨率下独立确定，与双域52-288结构中的核心域相同。
Insolubility of full-length HIV-1 integrase (IN) limited previous structure analyses to individual domains, By introducing five point mutations, we engineered a more soluble IN that allowed us to generate multidomain HIV-1 IN crystals. The first multidomain HIV-1 IN structure is reported. It incorporates the catalytic core and C-terminal domains (residues 52-288). The structure resolved to 2.8 Angstrom is a Y-shaped dimer, Within the dimer, the catalytic core domains form the only dimer interface, and the C-terminal domains are located 55 Angstrom apart, A 26-aa alpha-helix, alpha 6, links the C-terminal domain to the catalytic core. A kink in one of the two alpha 6 helices occurs near a known proteolytic site, suggesting that it may act as a flexible elbow to reorient the domains during the integration process. Two proteins that bind DNA in a sequence-independent manner are structurally homologous to the HIV-1 IN C-terminal domain, suggesting a similar protein-DNA interaction in which the IN C-terminal domain may serve to bind, bend, and orient viral DNA during integration, A strip of positively charged amino acids contributed by both monomers emerges from each active site of the dimer, suggesting a minimally dimeric platform for binding each viral DNA end. The crystal structure of the isolated catalytic core domain (residues 52-210), independently determined at 1.6-Angstrom resolution, is identical to the core domain within the two-domain 52-288 structure.