Allosteric Inhibition of Human Immunodeficiency Virus Integrase LATE BLOCK DURING VIRAL REPLICATION AND ABNORMAL MULTIMERIZATION INVOLVING SPECIFIC PROTEIN DOMAINS
Allosteric Inhibition of Human Immunodeficiency Virus Integrase LATE BLOCK DURING VIRAL REPLICATION AND ABNORMAL MULTIMERIZATION INVOLVING SPECIFIC PROTEIN DOMAINS
复制标题
DOI：
10.1074/jbc.m114.551119
复制
发表时间：
2014-07-25
影响因子：
4.8
通讯作者：
Bushman, Frederic D.
中科院分区：
文献类型：
--
作者：
Gupta, Kushol;Brady, Troy;Bushman, Frederic D.
HIV-1在抗病毒药物存在的情况下复制会导致耐药变异的进化，从而激励人们寻找更多的药物类别。在这里，我们报道了GSK1264的研究，它被鉴定为一种化合物，它破坏HIV-1整合酶(IN)和细胞因子晶状体上皮源性生长因子(LEDGF)/p75之间的相互作用。X-射线结晶学分析表明，GSK1264具有较强的抗病毒活性，其结合部位为LEDGF/p75 on IN。在GSK1264存在下对HIV复制的分析表明，只有轻微的抑制早期感染步骤，对整合靶向几乎没有影响，这是由LEDGF/p75.IN相互作用引导的。相反，对晚期复制步骤的抑制更有效。颗粒的产生是正常的，但颗粒的传染性降低了。GSK1264促进IN聚集，预制LEDGF/p75.IN复合体，提示其抑制机制。在聚集过程中，LEDGF/p75没有从IN移位，这表明LEDGF/p75被捕获在聚集体中。用截短的IN变异体进行的聚集分析表明，带有IN的催化结构域和C-末端结构域的结构只形成与药物诱导的有效聚集相关的开放聚合物。这些数据表明，IN的变构抑制剂是很有前途的抗病毒药物，并为其作用机制提供了新的信息。
HIV-1 replication in the presence of antiviral agents results in evolution of drug-resistant variants, motivating the search for additional drug classes. Here we report studies of GSK1264, which was identified as a compound that disrupts the interaction between HIV-1 integrase (IN) and the cellular factor lens epithelium-derived growth factor (LEDGF)/p75. GSK1264 displayed potent antiviral activity and was found to bind at the site occupied by LEDGF/p75 on IN by x-ray crystallography. Assays of HIV replication in the presence of GSK1264 showed only modest inhibition of the early infection steps and little effect on integration targeting, which is guided by the LEDGF/p75.IN interaction. In contrast, inhibition of late replication steps was more potent. Particle production was normal, but particles showed reduced infectivity. GSK1264 promoted aggregation of IN and preformed LEDGF/p75.IN complexes, suggesting a mechanism of inhibition. LEDGF/p75 was not displaced from IN during aggregation, indicating trapping of LEDGF/p75 in aggregates. Aggregation assays with truncated IN variants revealed that a construct with catalytic and C-terminal domains of IN only formed an open polymer associated with efficient drug-induced aggregation. These data suggest that the allosteric inhibitors of IN are promising antiviral agents and provide new information on their mechanism of action.