Potent nonnucleoside reverse transcriptase inhibitors target HIV-1 Gag-Pol.
Potent nonnucleoside reverse transcriptase inhibitors target HIV-1 Gag-Pol.
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DOI：
10.1371/journal.ppat.0020119
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发表时间：
2006-11
期刊：
影响因子：
6.7
通讯作者：
Tachedjian G
中科院分区：
文献类型：
--
作者：
Figueiredo A;Moore KL;Mak J;Sluis-Cremer N;de Bethune MP;Tachedjian G
非核苷类逆转录酶抑制剂（NNRTI）通过与HIV-1逆转录酶（RT）中接近但不同于DNA聚合酶活性位点的口袋结合来靶向RT，并阻止病毒cDNA的合成。NNRTI，特别是那些RT聚合酶活性的有效抑制剂，也可以作为酶的亚基间相互作用的化学增强剂。然而，这种化学增强作用对HIV-1复制的影响尚不清楚。在这里，我们表明，有效的非核苷类逆转录酶抑制剂依法韦仑，TMC120和TMC125，但不是奈韦拉平或地拉韦啶，抑制HIV-1复制的晚期阶段。这些有效的NNRTI增强了Gag和Gag-Pol多聚蛋白的细胞内加工，这与HIV-1转染细胞的病毒颗粒产生减少有关。增加的多蛋白加工与通过嵌入的RT序列通过NNRTI增强的Gag-Pol多聚化过早激活HIV-1蛋白酶一致。这些发现支持Gag-Pol多聚化是病毒组装中的重要步骤的观点，并证明Gag-Pol/Gag-Pol相互作用的调节是HIV-1产生的小分子抑制剂的新靶点。此外，这些药物可以作为有用的探针，以进一步了解参与HIV-1颗粒组装和成熟的过程。HIV-1编码逆转录酶（RT），这是一种病毒复制所必需的酶。非核苷类逆转录酶抑制剂（nonnucleotide reverse transcriptase inhibitors，NNRTI）是HIV-1逆转录酶的变构抑制剂。在HIV-1感染的细胞中，NNRTI阻断逆转录酶催化的病毒基因组RNA双链DNA拷贝的合成，这是病毒生命周期的早期步骤。有效的NNRTI具有促进两个RT亚基之间相互作用的新特征。然而，这种作用对抑制HIV-1复制的重要性尚未确定。在这项研究中，作者表明，有效的NNRTI阻断了病毒生命周期中的另一个步骤。非核苷类逆转录酶抑制剂增加了称为Gag和Gag-Pol的病毒多聚蛋白的细胞内加工，这些多聚蛋白表达HIV-1结构蛋白和病毒酶。增强的多蛋白加工与NNRTI处理细胞释放的病毒颗粒减少有关。NNRTI增强的多蛋白加工可能是由于药物与RT结合，表达为Gag-Pol多蛋白的一部分，并促进单独的Gag-Pol多蛋白之间的相互作用。这导致Gag-Pol嵌入的HIV-1蛋白酶的过早激活，导致可用于组装和从宿主细胞膜出芽的全长病毒多聚蛋白减少。这项研究提供了概念验证，即小分子可以调节Gag-Pol多聚蛋白之间的相互作用，并为HIV-1抗病毒药物的开发提供了新的靶点。
Nonnucleoside reverse transcriptase inhibitors (NNRTIs) target HIV-1 reverse transcriptase (RT) by binding to a pocket in RT that is close to, but distinct, from the DNA polymerase active site and prevent the synthesis of viral cDNA. NNRTIs, in particular, those that are potent inhibitors of RT polymerase activity, can also act as chemical enhancers of the enzyme's inter-subunit interactions. However, the consequences of this chemical enhancement effect on HIV-1 replication are not understood. Here, we show that the potent NNRTIs efavirenz, TMC120, and TMC125, but not nevirapine or delavirdine, inhibit the late stages of HIV-1 replication. These potent NNRTIs enhanced the intracellular processing of Gag and Gag-Pol polyproteins, and this was associated with a decrease in viral particle production from HIV-1-transfected cells. The increased polyprotein processing is consistent with premature activation of the HIV-1 protease by NNRTI-enhanced Gag-Pol multimerization through the embedded RT sequence. These findings support the view that Gag-Pol multimerization is an important step in viral assembly and demonstrate that regulation of Gag-Pol/Gag-Pol interactions is a novel target for small molecule inhibitors of HIV-1 production. Furthermore, these drugs can serve as useful probes to further understand processes involved in HIV-1 particle assembly and maturation. HIV-1 encodes reverse transcriptase (RT), an enzyme that is essential for virus replication. Nonnucleoside reverse transcriptase inhibitors (NNRTIs) are allosteric inhibitors of the HIV-1 RT. In HIV-1-infected cells NNRTIs block the RT-catalyzed synthesis of a double-stranded DNA copy of the viral genomic RNA, which is an early step in the virus life cycle. Potent NNRTIs have the novel feature of promoting the interaction between the two RT subunits. However, the importance of this effect on the inhibition of HIV-1 replication has not been defined. In this study, the authors show that potent NNRTIs block an additional step in the virus life cycle. NNRTIs increase the intracellular processing of viral polyproteins called Gag and Gag-Pol that express the HIV-1 structural proteins and viral enzymes. Enhanced polyprotein processing is associated with a decrease in viral particles released from NNRTI-treated cells. NNRTI enhanced polyprotein processing is likely due to the drug binding to RT, expressed as part of the Gag-Pol polyprotein and promoting the interaction between separate Gag-Pol polyproteins. This leads to premature activation of the Gag-Pol embedded HIV-1 protease, resulting in a decrease in full-length viral polyproteins available for assembly and budding from the host cell membrane. This study provides proof-of-concept that small molecules can modulate the interactions between Gag-Pol polyproteins and suggests a new target for the development of HIV-1 antiviral drugs.
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DOI：
10.1038/nsb0495-303
发表时间：
1995-04-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
ESNOUF, R;REN, JS;STUART, D
通讯作者：
STUART, D
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：
5.2
作者：
Almond, LM;Hoggard, PG;Back, DJ
通讯作者：
Back, DJ
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：
ADACHI, A;GENDELMAN, HE;MARTIN, MA
通讯作者：
MARTIN, MA