Small-Molecule Inhibition of Human Immunodeficiency Virus Type 1 Replication by Targeting the Interaction between Vif and ElonginC
Small-Molecule Inhibition of Human Immunodeficiency Virus Type 1 Replication by Targeting the Interaction between Vif and ElonginC
复制标题
通过靶向 Vif 和 ElonginC 之间的相互作用小分子抑制人类免疫缺陷病毒 1 型复制
DOI：
10.1128/jvi.06957-11
复制
发表时间：
2012-05-01
影响因子：
5.4
通讯作者：
Yu, Xianghui
中科院分区：
文献类型：
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作者：
Zuo, Tao;Liu, Donglai;Yu, Xianghui
HIV-1病毒感染因子（Vif）蛋白是病毒复制所必需的。Vif募集细胞ElonginB/C-Cullin 5 E3泛素连接酶以靶向宿主抗病毒蛋白APOBEC 3G（A3 G）进行蛋白酶体降解。在缺乏Vif的情况下，A3 G被包装到出芽的HIV-1病毒体中，并在新合成的负链病毒DNA中引入多个突变以限制病毒复制。因此，A3 G-Vif-E3复合物代表了用于开发新型抗HIV药物的有吸引力的靶标。在这项研究中，我们确定了一个有效的小分子化合物（VEC-5）的虚拟筛选和验证其抗Vif活性通过生化分析。我们发现，VEC-5抑制病毒复制仅在A3 G阳性细胞。当Vif共表达时，用VEC-5处理增加细胞A3 G水平，并增强A3 G掺入HIV-1病毒粒子以降低病毒感染性。免疫共沉淀和计算分析进一步将VEC-5的抗Vif活性归因于Vif与ElonginC蛋白直接结合的抑制。这些发现支持了抑制Vif功能可以释放A3 G以发挥其抗病毒活性的观点，并证明了Vif-ElonginC相互作用的调节是HIV-1小分子抑制剂的新靶点。
ABSTRACT The HIV-1 viral infectivity factor (Vif) protein is essential for viral replication. Vif recruits cellular ElonginB/C-Cullin5 E3 ubiquitin ligase to target the host antiviral protein APOBEC3G (A3G) for proteasomal degradation. In the absence of Vif, A3G is packaged into budding HIV-1 virions and introduces multiple mutations in the newly synthesized minus-strand viral DNA to restrict virus replication. Thus, the A3G-Vif-E3 complex represents an attractive target for development of novel anti-HIV drugs. In this study, we identified a potent small molecular compound (VEC-5) by virtual screening and validated its anti-Vif activity through biochemical analysis. We show that VEC-5 inhibits virus replication only in A3G-positive cells. Treatment with VEC-5 increased cellular A3G levels when Vif was coexpressed and enhanced A3G incorporation into HIV-1 virions to reduce viral infectivity. Coimmunoprecipitation and computational analysis further attributed the anti-Vif activity of VEC-5 to the inhibition of Vif from direct binding to the ElonginC protein. These findings support the notion that suppressing Vif function can liberate A3G to carry out its antiviral activity and demonstrate that regulation of the Vif-ElonginC interaction is a novel target for small-molecule inhibitors of HIV-1.