The y271 and i274 amino acids in reverse transcriptase of human immunodeficiency virus-1 are critical to protein stability.
The y271 and i274 amino acids in reverse transcriptase of human immunodeficiency virus-1 are critical to protein stability.
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DOI：
10.1371/journal.pone.0006108
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发表时间：
2009-07-03
期刊：
影响因子：
3.7
通讯作者：
Zheng BJ
中科院分区：
文献类型：
--
作者：
Zhang HJ;Wang YX;Wu H;Jin DY;Wen YM;Zheng BJ
人类免疫缺陷病毒（HIV）1的逆转录酶（RT）在启动病毒复制中起着关键作用，是开发抗HIV药物的重要靶点。我们的前期研究表明，逆转录酶（Gln 269-Arg 277）中的两个突变（Y271 A和I274 A）使病毒复制消失，但复制能力和逆转录酶活性不一致。在这项研究中，我们进一步研究了为什么在这两个位点的丙氨酸取代会影响病毒复制。我们发现，这两个突变体的病毒颗粒的RT活性和RT蛋白几乎检测不到，虽然Pr 160 gag-pol突变体的正确表达，运输和整合。利用蛋白酶抑制试验，我们证明了RT突变体的降解和病毒蛋白酶活性之间的相关性。我们的天然凝胶分析表明，在271和274个氨基酸的突变可能会导致构象变化，导致形成更高的顺序的寡聚体，而不是二聚体，导致增加蛋白质的不稳定性和对病毒蛋白酶的敏感性。因此，残基271和274对于RT稳定性和对病毒蛋白酶的抗性至关重要。这两个氨基酸残基在不同的HIV-1毒株中的保守性进一步支持了这一结论。这里获得的知识可能在药物设计中有用。
Reverse transcriptase (RT) of human immunodeficiency virus (HIV)-1 plays a key role in initiating viral replication and is an important target for developing anti-HIV drugs. Our previous study showed that two mutations (Y271A and I274A) in the turn RT (Gln269-Arg277) abrogated viral replication, but the replication capacity and RT activity was discordant. In this study, we further investigated why alanine substitutions at these two sites would affect viral replication. We found that both RT activity and RT protein were almost undetectable in viral particles of these two mutants, although the Pr160gag-pol mutants were properly expressed, transported and incorporated. Using protease inhibition assay, we demonstrated a correlation between the degradation of the RT mutants and the activity of viral protease. Our native gel analysis indicated that the mutations at 271 and 274 amino acids might cause conformational changes, leading to the formation of higher order oligomers instead of dimers, resulting in increased protein instability and susceptibility to viral protease. Thus, residues 271 and 274 are critical to RT stability and resistance to viral protease. The conservation of the two amino acid residues among different strains of HIV-1 lent further support to this conclusion. The knowledge gained here may prove useful in drug design.
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影响因子：
56.9
作者：
HARADA, S;KOYANAGI, Y;YAMAMOTO, N
通讯作者：
YAMAMOTO, N
DOI：
10.1073/pnas.90.13.6320
发表时间：
1993-07-01
影响因子：
11.1
作者：
JACOBOMOLINA, A;DING, JP;ARNOLD, E
通讯作者：
ARNOLD, E
影响因子：
5.4
作者：
Halwani, R;Khorchid, A;Kleiman, L
通讯作者：
Kleiman, L
影响因子：
5.4
作者：
PARK, J;MORROW, CD
通讯作者：
MORROW, CD
影响因子：
5.4
作者：
Abram, ME;Parniak, MA
通讯作者：
Parniak, MA