Structural Basis for Inhibitor-Induced Aggregation of HIV Integrase.
Structural Basis for Inhibitor-Induced Aggregation of HIV Integrase.
复制标题
DOI：
10.1371/journal.pbio.1002584
复制
发表时间：
2016-12
期刊：
影响因子：
9.8
通讯作者：
Bushman FD
中科院分区：
文献类型：
--
作者：
Gupta K;Turkki V;Sherrill-Mix S;Hwang Y;Eilers G;Taylor L;McDanal C;Wang P;Temelkoff D;Nolte RT;Velthuisen E;Jeffrey J;Van Duyne GD;Bushman FD
整合酶变构抑制剂（称为 ALLINI）通过与病毒编码的整合酶 (IN) 蛋白结合来干扰 HIV 复制。令人惊讶的是，ALLINI 不会干扰 DNA 整合，而是干扰 HIV 复制后期的病毒颗粒组装。为了研究 ALLINI 抑制机制，我们结晶了与 ALLINI GSK1264 结合的全长 HIV-1 IN，并以 4.4 Å 分辨率确定了复合物的结构。该结构显示 GSK1264 埋藏在 IN C 端结构域 (CTD) 和催化核心结构域之间。在晶格中，相互作用的域由两个不同的二聚体贡献，因此 IN 形成由抑制剂桥接接触介导的开放聚合物； N 端结构域不参与并且结构无序。抑制剂界面处的工程氨基酸取代阻断了 ALLINI 诱导的多聚化。对ALLINI敏感性降低的HIV逃逸突变体通常会改变抑制剂结合界面处或附近的氨基酸，并且这些取代也会减少IN多聚化。我们提出，ALLINI 通过催化核心结构域和 CTD 之间的相互作用刺激 IN 的不适当聚合来抑制颗粒组装，并且了解所涉及的界面为抑制剂优化提供了新途径。 HIV整合酶与变构抑制剂GSK1264复合物的新晶体结构解释了该药物如何诱导病毒蛋白聚集。一类有前途的新型抗病毒药物称为“ALLINI”（整合酶变构抑制剂），可有效抑制 HIV 复制。与其他药物一样，ALLINI 似乎也针对 HIV-1 整合酶 (IN)，这对该病毒的复制至关重要，但它们不是在 HIV 复制的早期阶段发挥作用，而是干扰晚期阶段发生的病毒颗粒组装和成熟，并诱导 IN 聚集。尽管有这些发现，但其影响的结构基础仍然未知。在这项研究中，我们结晶了全长 HIV-1 IN 与 ALLINI 的复合物（称为 GSK1264），并确定其结构为 4.4 Å。该结构首次揭示了完整的 ALLINI 结合界面，由 IN C 端和催化核心结构域组成。这些结构域由邻近的 IN 二聚体贡献，揭示了由抑制剂桥接接触介导的开放聚合构象。该界面处的取代阻止了 ALLINI 诱导的多聚化，并且我们发现针对此类药物的逃逸突变体位于该界面处或附近。我们提出 ALLINI 催化开放 IN 聚合物的形成，进而干扰病毒颗粒组装。
The allosteric inhibitors of integrase (termed ALLINIs) interfere with HIV replication by binding to the viral-encoded integrase (IN) protein. Surprisingly, ALLINIs interfere not with DNA integration but with viral particle assembly late during HIV replication. To investigate the ALLINI inhibitory mechanism, we crystallized full-length HIV-1 IN bound to the ALLINI GSK1264 and determined the structure of the complex at 4.4 Å resolution. The structure shows GSK1264 buried between the IN C-terminal domain (CTD) and the catalytic core domain. In the crystal lattice, the interacting domains are contributed by two different dimers so that IN forms an open polymer mediated by inhibitor-bridged contacts; the N-terminal domains do not participate and are structurally disordered. Engineered amino acid substitutions at the inhibitor interface blocked ALLINI-induced multimerization. HIV escape mutants with reduced sensitivity to ALLINIs commonly altered amino acids at or near the inhibitor-bound interface, and these substitutions also diminished IN multimerization. We propose that ALLINIs inhibit particle assembly by stimulating inappropriate polymerization of IN via interactions between the catalytic core domain and the CTD and that understanding the interface involved offers new routes to inhibitor optimization. A new crystal structure of the HIV integrase enzyme in complex with the allosteric inhibitor GSK1264 explains how the drug induces aggregation of the viral protein. A promising new class of antivirals called “ALLINIs” (allosteric inhibitors of integrase) potently inhibits HIV replication. Like other drugs, ALLINIs seem to target also the HIV-1 integrase (IN), which is crucial for the replication of this virus, but instead of acting at early phases of HIV replication, they interfere with viral particle assembly and maturation that occur at late stages and induce aggregation of IN. Despite these findings, the structural bases for the effects are still unknown. In this study, we crystallized full-length HIV-1 IN in complex with an ALLINI called GSK1264 and determined its structure to 4.4 Å. The structure reveals for the first time the complete ALLINI-binding interface, comprised of both IN C-terminal and catalytic core domains. These domains are contributed from neighboring IN dimers, revealing an open polymeric conformation mediated by inhibitor-bridged contacts. Substitutions at this interface block ALLINI-induced multimerization, and we find that escape mutants against this class of drug lie at or near this interface. We propose that ALLINIs catalyze formation of an open IN polymer, which in turn interferes with viral particle assembly.
登录
查看更多内容
影响因子：
5.7
作者：
Brunger, Axel T.;Adams, Paul D.;Fromme, Petra;Fromme, Raimund;Levitt, Michael;Schroeder, Gunnar F.
通讯作者：
Schroeder, Gunnar F.
影响因子：
5.4
作者：
Fontana, Juan;Jurado, Kellie A.;Steven, Alasdair C.
通讯作者：
Steven, Alasdair C.
影响因子：
2.9
作者：
Deprez, E;Tauc, P;Brochon, JC
通讯作者：
Brochon, JC
DOI：
10.1073/pnas.150220297
发表时间：
2000-07-18
影响因子：
11.1
作者：
Chen, JCH;Krucinski, J;Stroud, RM
通讯作者：
Stroud, RM
影响因子：
4.9
作者：
Fenwick, Craig;Amad, Ma'an;Yoakim, Christiane
通讯作者：
Yoakim, Christiane