HuR interacts with human immunodeficiency virus type 1 reverse transcriptase, and modulates reverse transcription in infected cells.
HuR interacts with human immunodeficiency virus type 1 reverse transcriptase, and modulates reverse transcription in infected cells.
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DOI：
10.1186/1742-4690-5-47
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发表时间：
2008-06-10
期刊：
影响因子：
3.3
通讯作者：
Liu LX
中科院分区：
文献类型：
--
作者：
Lemay J;Maidou-Peindara P;Bader T;Ennifar E;Rain JC;Benarous R;Liu LX
逆转录人类免疫缺陷病毒1型（HIV-1）的遗传物质是该病毒复制周期中的关键步骤。这一过程由逆转录酶（RT）催化，在生化水平上得到了很好的表征。然而，在受感染的细胞中，逆转录发生在多蛋白复合物中-逆转录复合物（RTC）-由与病毒蛋白（包括RT）相关的病毒基因组RNA组成，并且可能是尚未表征的细胞蛋白。关于与RTC相互作用的细胞蛋白质，特别是与逆转录酶相互作用的细胞蛋白质知之甚少。我们在这里报告说，HIV-1逆转录的影响水平的核质穿梭蛋白-RNA结合蛋白HuR。在酵母双杂交筛选中观察到RT和HuR之间的直接蛋白质-蛋白质相互作用，并通过均相时间分辨荧光（HTRF）在体外证实。我们将与HuR相互作用的结构域映射到RT的RNA酶H结构域，将RT的结合结构域映射到HuR的C末端，部分重叠HuR的第三个RRM RNA结合结构域。HuR沉默与特定的siRNA大大削弱了逆转录的早期和晚期步骤，显着抑制HIV-1感染。此外，通过诱变和免疫沉淀研究，我们不能检测到HuR与病毒RNA的结合。这些结果表明，HuR可能参与并可能调节HIV-1的逆转录反应，通过一种未知的机制，涉及与HIV-1 RT的蛋白质-蛋白质相互作用。
Reverse transcription of the genetic material of human immunodeficiency virus type 1 (HIV-1) is a critical step in the replication cycle of this virus. This process, catalyzed by reverse transcriptase (RT), is well characterized at the biochemical level. However, in infected cells, reverse transcription occurs in a multiprotein complex – the reverse transcription complex (RTC) – consisting of viral genomic RNA associated with viral proteins (including RT) and, presumably, as yet uncharacterized cellular proteins. Very little is known about the cellular proteins interacting with the RTC, and with reverse transcriptase in particular. We report here that HIV-1 reverse transcription is affected by the levels of a nucleocytoplasmic shuttling protein – the RNA-binding protein HuR. A direct protein-protein interaction between RT and HuR was observed in a yeast two-hybrid screen and confirmed in vitro by homogenous time-resolved fluorescence (HTRF). We mapped the domain interacting with HuR to the RNAse H domain of RT, and the binding domain for RT to the C-terminus of HuR, partially overlapping the third RRM RNA-binding domain of HuR. HuR silencing with specific siRNAs greatly impaired early and late steps of reverse transcription, significantly inhibiting HIV-1 infection. Moreover, by mutagenesis and immunoprecipitation studies, we could not detect the binding of HuR to the viral RNA. These results suggest that HuR may be involved in and may modulate the reverse transcription reaction of HIV-1, by an as yet unknown mechanism involving a protein-protein interaction with HIV-1 RT.
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影响因子：
5.4
作者：
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通讯作者：
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影响因子：
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影响因子：
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影响因子：
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影响因子：
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作者：
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