MiniCD4 protein resistance mutations affect binding to the HIV-1 gp120 CD4 binding site and decrease entry efficiency
MiniCD4 protein resistance mutations affect binding to the HIV-1 gp120 CD4 binding site and decrease entry efficiency
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DOI：
10.1186/1742-4690-9-36
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发表时间：
2012-05-02
期刊：
影响因子：
3.3
通讯作者：
Arien, Kevin K.
中科院分区：
文献类型：
--
作者：
Grupping, Katrijn;Selhorst, Philippe;Arien, Kevin K.
背景：病毒包膜蛋白 (Env)，特别是其 gp120 亚基与细胞 CD4 受体的结合是 HIV-1 进入过程的第一个重要步骤。已知 gp120 的 CD4 结合位点 (CD4bs)，特别是由 CD4 Phe43 残基占据的凹腔，在不同的循环亚型中高度保守，因此构成了疫苗和药物设计特别有趣的目标。 miniCD4 蛋白是一类有前途的 CD4bs 抑制剂。研究 CD4bs 抑制剂压力下的病毒进化可以深入了解 gp120-CD4 相互作用和病毒进入。结果：本研究报告了两种 B 亚型 HIV-1 对最活跃的 miniCD4 M48U1 及其祖先 M48 的耐药诱导，以及这些突变位置如何影响 CD4bs 识别、进入效率和对其他 CD4bs 抑制剂的敏感性。 BaL 和 SF162 中对 M48U1 的耐药性始终与 S375R/N 取代有关； M48 抗性与 SF162 中的 D474N 取代和 BaL 中的 H105Y 取代相关。此外，V255 和 G471 位点的一些其他突变对于 SF162 抗性病毒也很重要。除474外，所有这些突变位置都是保守的，将它们引入表达SF162 Env的感染性分子克隆（pBRNL4.3 SF162）中会导致进入效率降低。此外，耐药突变体至少对其他CD4bs抑制剂、V3单克隆抗体447-52D表现出一定的交叉耐药性，有些甚至对单克隆抗体17b表现出交叉耐药性，其表位与共受体结合位点重叠。结论：发现突变H105Y、V255M、S375R/N、G471R/E和D474N与对CD4bs抑制剂的耐药性有关。 M48 和 M48U1。所有突变位置都是 CD4b 的一部分或与其非常接近；大多数都是高度保守的，并且都会对进入效率产生影响，这表明它们对于最佳病毒感染性的重要性。
Background: Binding of the viral envelope protein (Env), and particularly of its gp120 subunit, to the cellular CD4 receptor is the first essential step of the HIV-1 entry process. The CD4 binding site ( CD4bs) of gp120, and especially a recessed cavity occupied by the CD4 Phe43 residue, are known to be highly conserved among the different circulating subtypes and therefore constitute particularly interesting targets for vaccine and drug design. The miniCD4 proteins are a promising class of CD4bs inhibitors. Studying virus evolution under pressure of CD4bs inhibitors could provide insight on the gp120-CD4 interaction and viral entry.Results: The present study reports on the resistance induction of two subtype B HIV-1 against the most active miniCD4, M48U1, and its ancestor, M48, and how these mutated positions affect CD4bs recognition, entry efficiency, and sensitivity to other CD4bs inhibitors. Resistance against M48U1 was always associated with S375R/N substitution in both BaL and SF162; M48 resistance was associated with D474N substitution in SF162 and with H105Y substitution in BaL. In addition, some other mutations at position V255 and G471 were of importance for SF162 resistant viruses. Except for 474, all of these mutated positions are conserved, and introducing them into an SF162 Env expressing infectious molecular clone ( pBRNL4.3 SF162) resulted in decreased entry efficiency. Furthermore, resistant mutants showed at least some cross-resistance towards other CD4bs inhibitors, the V3 monoclonal antibody 447-52D and some even against the monoclonal antibody 17b, of which the epitope overlaps the co-receptor binding site.Conclusions: The mutations H105Y, V255M, S375R/N, G471R/E, and D474N are found to be involved in resistance towards M48 and M48U1. All mutated positions are part of, or in close proximity to, the CD4bs; most are highly conserved, and all have an impact on the entry efficiency, suggesting their importance for optimal virus infectivity.