Structural basis for the role of the K65R mutation in HIV-1 reverse transcriptase polymerization, excision antagonism, and tenofovir resistance.
Structural basis for the role of the K65R mutation in HIV-1 reverse transcriptase polymerization, excision antagonism, and tenofovir resistance.
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K65R突变在HIV-1逆转录酶聚合，切除拮抗和替诺福韦抗性中的作用的结构基础。
DOI：
10.1074/jbc.m109.022525
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发表时间：
2009-12-11
期刊：
影响因子：
--
通讯作者：
Arnold E
中科院分区：
文献类型：
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作者：
Das K;Bandwar RP;White KL;Feng JY;Sarafianos SG;Tuske S;Tu X;Clark AD Jr;Boyer PL;Hou X;Gaffney BL;Jones RA;Miller MD;Hughes SH;Arnold E
K65R是一种主要的逆转录酶(RT)突变，选择在人类免疫缺陷病毒1型感染患者中使用含有替诺福韦、富马酸二异丙酯或其他核苷类似RT药物的抗逆转录病毒方案。我们测定了K65R突变体RT与双链DNA以及与替诺福韦二磷酸(TFV-DP)或dATP的复合物中的晶体结构。晶体允许在dNTP结合部位用dATP取代TFV-DP。精氨酸K65R和Arg72的胍基平面被堆积形成一个分子平台，限制了这两个残基的构象适应性，这解释了K65R突变对核苷酸掺入和切除的负面影响。此外，K65R和Arg72的胍平面在TFV-dp和dATP结合结构中以两种不同的旋转体构象堆积，这可能有助于解释K65R RT如何区分药物和底物。K65R介导的这些对RT结构和功能的影响有助于我们可视化与其他关键核苷酸RT耐药突变的复杂相互作用，如M184V、L74V和胸苷类似物耐药突变。
K65R is a primary reverse transcriptase (RT) mutation selected in human immunodeficiency virus type 1-infected patients taking antiretroviral regimens containing tenofovir disoproxil fumarate or other nucleoside analog RT drugs. We determined the crystal structures of K65R mutant RT cross-linked to double-stranded DNA and in complexes with tenofovir diphosphate (TFV-DP) or dATP. The crystals permit substitution of TFV-DP with dATP at the dNTP-binding site. The guanidinium planes of the arginines K65R and Arg72 were stacked to form a molecular platform that restricts the conformational adaptability of both of the residues, which explains the negative effects of the K65R mutation on nucleotide incorporation and on excision. Furthermore, the guanidinium planes of K65R and Arg72 were stacked in two different rotameric conformations in TFV-DP- and dATP-bound structures that may help explain how K65R RT discriminates the drug from substrates. These K65R-mediated effects on RT structure and function help us to visualize the complex interaction with other key nucleotide RT drug resistance mutations, such as M184V, L74V, and thymidine analog resistance mutations.