Essential regions of the tRNA primer required for HIV-1 infectivity.
Essential regions of the tRNA primer required for HIV-1 infectivity.
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DOI：
10.1093/nar/28.23.4783
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发表时间：
2000-12
影响因子：
14.9
通讯作者：
Q. Yu;C. Morrow
中科院分区：
文献类型：
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作者：
Q. Yu;C. Morrow
人类免疫缺陷病毒(HIV)像所有的逆转录病毒一样，需要细胞内的tRNA作为启动逆转录的引子。在以前的研究中，我们证明了带有与酵母tRNA(Phe)互补的引物结合位点的HIV-1(psHIV-Phe)除非酵母tRNA(Phe)以反式形式提供，否则不具有传染性。这一独特的体内互补系统现已被用于确定HIV-1复制所需的tRNA元件。构建了tRNA的TPsiC茎环、反密码子茎环或D茎环缺失的突变型tRNA(Phe)，并对其拯救pHIV-Phe的能力进行了评估。带有中断的TPsiC茎环的突变体tRNA(Phe)不能挽救pHIV-Phe。相比之下，没有D茎环的突变体tRNA(Phe)完全可以用于救援。研究发现，tRNA反密码子茎环区域对于有效互补是重要的。我们的研究结果首次证明了tRNA引物的特定结构和序列元件对HIV-1反转录的重要性，并为阻断HIV-1复制确定了新的靶点。
Human immunodeficiency virus (HIV), like all retroviruses, requires a cellular tRNA as a primer for initiation of reverse transcription. In a previous study, we demonstrated that an HIV-1 with a primer binding site complementary to yeast tRNA(Phe) (psHIV-Phe) was not infectious unless yeast tRNA(Phe) was supplied in trans. This unique in vivo complementation system has now been used to define the elements of the tRNA required for HIV-1 replication. Mutant tRNA(Phe) with deletions in TPsiC stem-loop, anticodon stem-loop or D stem-loop of the tRNA were generated and assessed for the capacity to rescue psHIV-Phe. Mutant tRNA(Phe) with disrupted TPsiC stem-loop did not rescue psHIV-Phe. In contrast, a mutant tRNA(Phe) without the D stem-loop was fully functional for the rescue. The tRNA anticodon stem-loop region was found to be important for efficient complementation. The results of our studies demonstrate for the first time the importance of specific structural and sequence elements of the tRNA primer for HIV-1 reverse transcription and define new targets for interruption of HIV-1 replication.