The Effect of Clade-Specific Sequence Polymorphisms on HIV-1 Protease Activity and Inhibitor Resistance Pathways
The Effect of Clade-Specific Sequence Polymorphisms on HIV-1 Protease Activity and Inhibitor Resistance Pathways
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DOI：
10.1128/jvi.00505-10
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发表时间：
2010-10-01
影响因子：
5.4
通讯作者：
Schiffer, Celia A.
中科院分区：
文献类型：
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作者：
Bandaranayake, Rajintha M.;Kolli, Madhavi;Schiffer, Celia A.
全世界大多数 HIV-1 感染都是由非 B 进化枝 HIV-1 毒株引起的。 CRF01_AE (AE) 菌株主要见于东南亚。 AE 蛋白酶与进化枝 B 蛋白酶的氨基酸序列相差约 10%，并且携带一些与进化枝 B 中的耐药性相关的天然存在的多态性。已观察到 AE 蛋白酶通过奈非那韦 (NFV) 治疗的非活性位点 N88S 突变产生耐药性，而进化枝 B 蛋白酶同时产生活性位点突变 D30N 和非活性位点突变 N88D。使用野生型和 NFV 抗性进化枝 B 和 AE 蛋白酶变体进行结构和生化研究。还评估了进化枝特异性序列变异与抑制剂抗性途径之间的关系。 AE 蛋白酶的催化周转率比进化枝 B 蛋白酶低，并且对 NFV 和地芦那韦 (DRV) 的亲和力也较弱。这种较弱的亲和力可能导致 AE 中的非活性位点 N88S 变体，其对 NFV 和 DRV 的亲和力显着降低。 B 进化枝中的 D30N/N88D 突变导致对 NFV 的亲和力显着丧失，并且在较小程度上导致对 DRV 的亲和力丧失。 AE 蛋白酶晶体结构的比较显示，与进化枝 B 蛋白酶相比，瓣铰链区发生了显着的结构重排，并提出了对 NFV 抗性的替代途径的见解。综上所述，我们的研究表明，进化枝内的序列多态性可以改变蛋白酶活性和抑制剂结合，并且能够改变抑制剂抗性的途径。
The majority of HIV-1 infections around the world result from non-B clade HIV-1 strains. The CRF01_AE (AE) strain is seen principally in Southeast Asia. AE protease differs by similar to 10% in amino acid sequence from clade B protease and carries several naturally occurring polymorphisms that are associated with drug resistance in clade B. AE protease has been observed to develop resistance through a nonactive-site N88S mutation in response to nelfinavir (NFV) therapy, whereas clade B protease develops both the active-site mutation D30N and the nonactive-site mutation N88D. Structural and biochemical studies were carried out with wild-type and NFV-resistant clade B and AE protease variants. The relationship between clade-specific sequence variations and pathways to inhibitor resistance was also assessed. AE protease has a lower catalytic turnover rate than clade B protease, and it also has weaker affinity for both NFV and darunavir (DRV). This weaker affinity may lead to the nonactive-site N88S variant in AE, which exhibits significantly decreased affinity for both NFV and DRV. The D30N/N88D mutations in clade B resulted in a significant loss of affinity for NFV and, to a lesser extent, for DRV. A comparison of crystal structures of AE protease shows significant structural rearrangement in the flap hinge region compared with those of clade B protease and suggests insights into the alternative pathways to NFV resistance. In combination, our studies show that sequence polymorphisms within clades can alter protease activity and inhibitor binding and are capable of altering the pathway to inhibitor resistance.