A Conformational Escape Reaction of HIV-1 against an Allosteric Integrase Inhibitor
A Conformational Escape Reaction of HIV-1 against an Allosteric Integrase Inhibitor
复制标题
DOI：
10.1128/jvi.00486-20
复制
发表时间：
2020-10-01
影响因子：
5.4
通讯作者：
Nakata，Hirotomo
中科院分区：
文献类型：
--
作者：
Nakamura，Tomofumi;Nakamura，Teruya;Nakata，Hirotomo
尽管抗逆转录病毒治疗（ART）有好处，但HIV-1经常获得耐药突变。HIV-1整合酶（IN）对于HIV-1 DNA整合到宿主基因组中至关重要。IN进一步促进HIV-1 RNA结合，这是HIV-1成熟所必需的。非催化位点整合酶抑制剂（NCINIs）已被开发为变抗性IN抑制剂，其通过多模式的作用发挥抗HIV-1活性，如在早期抑制IN晶状体上皮衍生生长因子(LEDGF)/p75相互作用，在HIV-1复制后期破坏功能性IN多聚。在这里，我们发现IN经历了一个适应性的构象变化来逃避ncini。我们观察到耐ncni的HIV-1变异在in编码区通过26代（P26）积累了4个氨基酸突变。我们利用高效液相色谱（HPLC）、热稳定性分析和x射线晶体学分析表明，一些氨基酸突变会影响IN催化核心结构域（CCDs）的稳定性和/或二聚化界面，可能导致全长IN蛋白的多聚性严重降低（IN的多聚性不足）。这种通过ncni相关突变的低聚合的IN被HIV-1 RNA稳定，并恢复到病毒颗粒中与野生型HIV-1相同的水平。重组HIV-1克隆与野生型HIV-1相似。我们的研究表明，作为逃逸机制之一，HIV-1最终可以通过IN过聚合来抵消ncini诱导的IN过聚合。我们的研究结果为理解带有或不带有HIV-1 RNA的IN多聚提供了信息，并可能影响抗HIV-1策略的发展。了解HIV-1对抗HIV-1药物的耐药机制可能导致开发效率更高的新药，从而产生更有效的抗逆转录病毒治疗。由更有效和长效抗HIV-1药物组成的抗逆转录病毒疗法可以大大提高药物依从性，并提供HIV-1预防，如暴露前预防。具有多模态作用模式的NCINIs在HIV-1成熟过程中通过IN过聚合发挥了有效的抗HIV-1作用。然而，HIV-1可以获得一些导致IN低聚合的突变，以减轻ncni诱导的IN过聚合。这种低聚合的IN被HIV-1 RNA有效地稳定，并恢复到与野生型HIV-1相同的水平。我们的研究结果表明，HIV-1最终获得了这种构象逃逸反应来克服独特的NCINI作用。研究与HIV-1蛋白多聚相关的耐药突变可能有助于阐明其分子机制和功能多聚，使我们能够开发更有效的抗HIV-1药物和独特的治疗策略。
HIV-1 often acquires drug-resistant mutations in spite of the benefits of antiretroviral therapy (ART). HIV-1 integrase (IN) is essential for the concerted integration of HIV-1 DNA into the host genome. IN further contributes to HIV-1 RNA binding, which is required for HIV-1 maturation. Non-catalytic-site integrase inhibitors (NCINIs) have been developed as allosteric IN inhibitors, which perform anti-HIV-1 activity by a multimodal mode of action such as inhibition of the IN-lens epithelium-derived growth factor (LEDGF)/p75 interaction in the early stage and disruption of functional IN multimerization in the late stage of HIV-1 replication. Here, we show that IN undergoes an adaptable conformational change to escape from NCINIs. We observed that NCINI-resistant HIV-1 variants have accumulated 4 amino acid mutations by passage 26 (P26) in the IN-encoding region. We employed high-performance liquid chromatography (HPLC), thermal stability assays, and X-ray crystallographic analysis to show that some amino acid mutations affect the stability and/or dimerization interface of the IN catalytic core domains (CCDs), potentially resulting in the severely decreased multimerization of full-length IN proteins (IN undermultimerization). This undermultimerized IN via NCINI-related mutations was stabilized by HIV-1 RNA and restored to the same level as that of wild-type HIV-1 in viral particles. Recombinant HIV-1 clones with IN undermultimerization propagated similarly to wild-type HIV-1. Our study revealed that HIV-1 can eventually counteract NCINI-induced IN overmultimerization by IN undermultimerization as one of the escape mechanisms. Our findings provide information on the understanding of IN multimerization with or without HIV-1 RNA and may influence the development of anti-HIV-1 strategies.IMPORTANCEUnderstanding the mechanism of HIV-1 resistance to anti-HIV-1 drugs could lead to the development of novel drugs with increased efficiency, resulting in more effective ART. ART composed of more potent and long-acting anti-HIV-1 drugs can greatly improve drug adherence and also provide HIV-1 prevention such as preexposure prophylaxis. NCINIs with a multimodal mode of action exert potent anti-HIV-1 effects through IN overmultimerization during HIV-1 maturation. However, HIV-1 can acquire some mutations that cause IN undermultimerization to alleviate NCINI-induced IN overmultimerization. This undermultimerized IN was efficiently stabilized by HIV-1 RNA and restored to the same level as that of wild-type HIV-1. Our findings revealed that HIV-1 eventually acquires such a conformational escape reaction to overcome the unique NCINI actions. The investigation into drug-resistant mutations associated with HIV-1 protein multimerization may facilitate the elucidation of its molecular mechanism and functional multimerization, allowing us to develop more potent anti-HIV-1 drugs and unique treatment strategies.