Nucleotides within the anticodon stem are important for optimal use of tRNA(Lys,3) as the primer for HIV-1 reverse transcription.

Nucleotides within the anticodon stem are important for optimal use of tRNA(Lys,3) as the primer for HIV-1 reverse transcription.
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反密码子茎内的核苷酸对于最佳使用 tRNA(Lys,3) 作为 HIV-1 逆转录引物非常重要。

DOI:
10.1016/j.virol.2007.02.010
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发表时间:
2007
期刊:
影响因子:
3.7
通讯作者:
Morrow,CaseyD
Morrow,CaseyD
中科院分区:
医学3区
文献类型:
--
作者:
McCulley,Anna;Morrow,CaseyD

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HIV-1利用tRNALys,3作为逆转录起始的引物。为了进一步研究tRNA序列和引物选择的结构要求,我们开发了一个互补系统,该系统需要以反式提供tRNALys。我们构建了一个HIV-1前病毒,其中引物结合位点(PBS)被改变为与E. coli tRNALys,3与哺乳动物tRNALys,3有许多相同的碱基,证明只有将原病毒与编码大肠杆菌tRNALys,3的质粒共转染才能获得感染性病毒。colitRNALys3.在目前的研究中,我们突变了E。coli tRNALys,3,使反密码子茎环的茎内核苷酸与哺乳动物tRNALys,3相同。互补性分析表明,改造后的E. coli tRNALys,3(E. coli tRNALys,3-MA)的互补效率是E. colitRNALys3.在E. colitRNALys,3-MA不同于E. colitRNALys,3揭示了核苷酸序列在逆转录中的重要性。我们的研究结果强调了哺乳动物tRNALys,3的多个区域对于tRNALys,3作为HIV-1逆转录的引物的偏好是重要的。
HIV-1 utilizes tRNALys,3as the primer for initiation of reverse transcription. To further examine the tRNA sequence and structural requirements for primer selection, we developed a complementation system which required tRNALysto be provided in trans. We constructed an HIV-1 provirus in which the primer-binding site (PBS) was altered to be complementary to the 3′ terminal 18-nucleotides of E. coli tRNALys,3, which shares many bases with mammalian tRNALys,3, and demonstrated that infectious virus was obtained only if the provirus was co-transfected with the plasmid encoding E. coli tRNALys,3. In the current study we have mutated E. coli tRNALys,3so that nucleotides within the stem of the anticodon stem–loop were made identical to mammalian tRNALys,3. Analysis of the complementation revealed that the modified E. coli tRNALys,3(E. coli tRNALys,3-MA) complemented 3–5 times more efficiently than E. coli tRNALys,3. Mutation of nucleotides within the anticodon stem region of E. coli tRNALys,3-MA that differed from E. coli tRNALys,3revealed the importance of the nucleotide sequence for efficient use in reverse transcription. The results of our studies highlight that multiple regions of mammalian tRNALys,3are important for the preference of tRNALys,3as the primer for HIV-1 reverse transcription.
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