分析 HIV-1 中的非赖氨酰 tRNA。
Profiling non-lysyl tRNAs in HIV-1.
出版信息
RNA. 2010 Feb;16(2):267-73. doi: 10.1261/rna.1928110. Epub 2009 Dec 9.
Abstract
During its assembly, human HIV-1 selectively packages the tRNA(Lys) isoacceptors, including tRNA(Lys3), the primer for the reverse transcriptase. However, other low molecular weight RNA species are also seen in the virus. We profiled the tRNAs packaged into HIV-1 using microarray analysis and validated our results by two-dimensional gel electrophoresis and RT-PCR. In addition to tRNA(Lys) isoacceptors, tRNA(Asn) and the rare isoacceptor of tRNA(Ile) are also selectively packaged. In Gag viral-like particles missing the GagPol protein, overall tRNA incorporation is reduced by >80%. This reduction is significantly greater than can be accounted for by the reduction in tRNA(Lys) isoacceptors, tRNA(Asn) and tRNA(Ile), suggesting that incorporation of other tRNAs may also require the GagPol protein. These results demonstrate selective incorporation of non-lysyl tRNAs into HIV-1 and highlight the application of microarrays as a novel method to study tRNA incorporation into viruses.
摘要
在组装过程中，人类 HIV-1 选择性地包装 tRNA(Lys)同功受体，包括 tRNA(Lys3)，这是逆转录酶的引物。然而，病毒中也可以看到其他低分子量 RNA 种类。我们使用微阵列分析对包装到 HIV-1 中的 tRNAs 进行了分析，并通过二维凝胶电泳和 RT-PCR 验证了我们的结果。除了 tRNA(Lys)同功受体外，tRNA(Asn)和 tRNA(Ile)的稀有同功受体也被选择性包装。在缺乏 GagPol 蛋白的 Gag 病毒样颗粒中，总的 tRNA 掺入减少了>80%。这种减少明显大于 tRNA(Lys)同功受体、tRNA(Asn)和 tRNA(Ile)减少的程度，这表明其他 tRNA 的掺入也可能需要 GagPol 蛋白。这些结果表明非赖氨酸 tRNA 被选择性地掺入 HIV-1 中，并强调了微阵列作为一种研究 tRNA 掺入病毒的新方法的应用。
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