Glyceraldehyde 3-phosphate dehydrogenase negatively regulates human immunodeficiency virus type 1 infection.
Glyceraldehyde 3-phosphate dehydrogenase negatively regulates human immunodeficiency virus type 1 infection.
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DOI：
10.1186/1742-4690-9-107
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发表时间：
2012-12-13
期刊：
影响因子：
3.3
通讯作者：
Misumi S
中科院分区：
文献类型：
--
作者：
Kishimoto N;Onitsuka A;Kido K;Takamune N;Shoji S;Misumi S
宿主蛋白在装配过程中被整合到人类免疫缺陷病毒1型（HIV-1）病毒粒子内，并可以正向或负向调节HIV-1感染。虽然质谱法提高了宿主蛋白的鉴定效率，但这些宿主蛋白如何影响HIV-1复制尚未完全阐明。在这项研究中，我们表明，病毒体相关的甘油醛3-磷酸脱氢酶（GAPDH）不变构的HIV-1逆转录酶（RT），但降低逆转录反应的效率，通过降低包装效率赖氨酰-tRNA合成酶（LysRS）和tRNALys 3到HIV-1病毒体。双向凝胶电泳结果表明，在产HIV-1的CEM/LAV-1细胞中，GAPDH的一些同工酶具有不同的等电点，并有一定比例的GAPDH被选择性地掺入到病毒颗粒中。在CEM/LAV-1细胞中通过RNA干扰抑制GAPDH表达导致病毒粒子内GAPDH包装减少，并且GAPDH包装缺陷型病毒保持至少控制水平的病毒生产，但增加感染性。逆转录产物的定量分析表明，GAPDH包装缺陷型病毒的早期cDNA产物的水平高于对照病毒的水平，这是由于LysRS和tRNALys 3进入病毒体的包装效率较高，而不是GAPDH依赖性的负变构调节RT。使用抗GAPDH抗体的免疫沉淀分析显示GAPDH直接与Pr 55 gag和p160 gag-pol相互作用，并且LysRS在HIV-1中过表达。生产细胞导致GAPDH包装在HIV颗粒中的效率降低。相比之下，由表达高水平GAPDH的细胞产生的病毒在TZM-bl细胞中显示出降低的感染性，并且在TZM-bl细胞和外周血单核细胞（PBMC）中显示出降低的逆转录效率。这些发现表明GAPDH负调控HIV-1感染，并为GAPDH在HIV-1生命周期中的新功能和抗HIV-1感染的新宿主防御机制提供了见解。
Host proteins are incorporated inside human immunodeficiency virus type 1 (HIV-1) virions during assembly and can either positively or negatively regulate HIV-1 infection. Although the identification efficiency of host proteins is improved by mass spectrometry, how those host proteins affect HIV-1 replication has not yet been fully clarified. In this study, we show that virion-associated glyceraldehyde 3-phosphate dehydrogenase (GAPDH) does not allosterically inactivate HIV-1 reverse transcriptase (RT) but decreases the efficiency of reverse transcription reactions by decreasing the packaging efficiency of lysyl-tRNA synthetase (LysRS) and tRNALys3 into HIV-1 virions. Two-dimensional (2D) gel electrophoresis demonstrated that some isozymes of GAPDH with different isoelectric points were expressed in HIV-1-producing CEM/LAV-1 cells, and a proportion of GAPDH was selectively incorporated into the virions. Suppression of GAPDH expression by RNA interference in CEM/LAV-1 cells resulted in decreased GAPDH packaging inside the virions, and the GAPDH-packaging-defective virus maintained at least control levels of viral production but increased the infectivity. Quantitative analysis of reverse transcription products indicated that the levels of early cDNA products of the GAPDH-packaging-defective virus were higher than those of the control virus owing to the higher packaging efficiencies of LysRS and tRNALys3 into the virions rather than the GAPDH-dependent negative allosteric modulation for RT. Furthermore, immunoprecipitation assay using an anti-GAPDH antibody showed that GAPDH directly interacted with Pr55gag and p160gag-pol and the overexpression of LysRS in HIV-1-producing cells resulted in a decrease in the efficiency of GAPDH packaging in HIV particles. In contrast, the viruses produced from cells expressing a high level of GAPDH showed decreased infectivity in TZM-bl cells and reverse transcription efficiency in TZM-bl cells and peripheral blood mononuclear cells (PBMCs). These findings indicate that GAPDH negatively regulates HIV-1 infection and provide insights into a novel function of GAPDH in the HIV-1 life cycle and a new host defense mechanism against HIV-1 infection.
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DOI：
10.1073/pnas.222542999
发表时间：
2002-10-29
影响因子：
11.1
作者：
Glaser, PE;Han, XL;Gross, RW
通讯作者：
Gross, RW
影响因子：
3.8
作者：
Petrik, J;Parker, H;Alexander, GJM
通讯作者：
Alexander, GJM
影响因子：
5.4
作者：
Ikeda, T;Nishitsuji, H;Masuda, T
通讯作者：
Masuda, T
DOI：
10.1073/pnas.93.18.9937
发表时间：
1996-09-03
影响因子：
11.1
作者：
Ishitani, R;Chuang, DM
通讯作者：
Chuang, DM
影响因子：
3.7
作者：
Ott, DE;Coren, LV;Henderson, LE
通讯作者：
Henderson, LE