Placement of leucine zipper motifs at the carboxyl terminus of HIV-1 protease significantly reduces virion production.
Placement of leucine zipper motifs at the carboxyl terminus of HIV-1 protease significantly reduces virion production.
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DOI：
10.1371/journal.pone.0032845
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Wang CT
中科院分区：
文献类型：
--
作者：
Pan YY;Wang SM;Huang KJ;Chiang CC;Wang CT
天然HIV-1蛋白酶（PR）是同源二聚体。一些研究人员认为，HIV-1 Gag-Pol分子之间的相互作用触发了嵌入式PR的激活（介导Gag和Gag-Pol切割），PR外的Gag-Pol组装结构域可能通过影响Gag-Pol背景下的PR二聚体相互作用而促进PR激活。为了确定PR二聚体相互作用的增强是否促进PR活化，我们将单个或串联重复亮氨酸拉链（LZ）置于PR C末端，并寻找增强的Gag加工效率和增加的Gag-PR-LZ多聚化能力之间的相关性。我们发现HIV-1突变体产生的病毒样颗粒（VLP）显著减少，由于Gag切割效率增强，LZ融合到PR的末端。由于在PR活性抑制后VLP产生可以恢复到wt水平，因此认为该组装缺陷是PR活性依赖性的。我们还发现LZ增强对Gag切割的作用与增强的Gag-PR多聚化之间存在相关性。结果表明，PR二聚体的相互作用促进强制Gag-PR多聚化导致过早的Gag裂解，可能是过早的PR激活的结果。我们的结论是，PR下游的异源二聚化结构域的放置增强PR介导的Gag切割效率，这意味着结构构象，而不是PR外的一级序列，是HIV-1 PR激活的主要决定因素。
Natural HIV-1 protease (PR) is homodimeric. Some researchers believe that interactions between HIV-1 Gag-Pol molecules trigger the activation of embedded PR (which mediates Gag and Gag-Pol cleavage), and that Gag-Pol assembly domains outside of PR may contribute to PR activation by influencing PR dimer interaction in a Gag-Pol context. To determine if the enhancement of PR dimer interaction facilitates PR activation, we placed single or tandem repeat leucine zippers (LZ) at the PR C-terminus, and looked for a correlation between enhanced Gag processing efficiency and increased Gag-PR-LZ multimerization capacity. We found significant reductions in virus-like particles (VLPs) produced by HIV-1 mutants, with LZ fused to the end of PR as a result of enhanced Gag cleavage efficiency. Since VLP production can be restored to wt levels following PR activity inhibition, this assembly defect is considered PR activity-dependent. We also found a correlation between the LZ enhancement effect on Gag cleavage and enhanced Gag-PR multimerization. The results suggest that PR dimer interactions facilitated by forced Gag-PR multimerization lead to premature Gag cleavage, likely a result of premature PR activation. Our conclusion is that placement of a heterologous dimerization domain downstream of PR enhances PR-mediated Gag cleavage efficiency, implying that structural conformation, rather than the primary sequence outside of PR, is a major determinant of HIV-1 PR activation.
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影响因子：
6.7
作者：
Figueiredo A;Moore KL;Mak J;Sluis-Cremer N;de Bethune MP;Tachedjian G
通讯作者：
Tachedjian G
影响因子：
5.4
作者：
ARRIGO, SJ;HUFFMAN, K
通讯作者：
HUFFMAN, K
DOI：
10.1073/pnas.86.15.5781
发表时间：
1989-08-01
影响因子：
11.1
作者：
GOTTLINGER, HG;SODROSKI, JG;HASELTINE, WA
通讯作者：
HASELTINE, WA
影响因子：
3.7
作者：
Liao, WH;Wang, CT
通讯作者：
Wang, CT
影响因子：
5.4
作者：
Chiu, HC;Yao, SY;Wang, CT
通讯作者：
Wang, CT