HIV-1 RNA Trafficking Laboratory; Lady Davis Institute at the Jewish General Hospital; Montréal, Québec, Canada; Department of Microbiology & Immunology; McGill University; Montréal, Québec, Canada.
HIV-1 RNA Trafficking Laboratory; Lady Davis Institute at the Jewish General Hospital; Montréal, Québec, Canada; Department of Medicine; Division of Experimental Medicine; McGill University; Montréal, Québec, Canada.
RNA Biol. 2013 Nov;10(11):1714-25. doi: 10.4161/rna.26542. Epub 2013 Sep 30.
hnRNP A2 is a cellular protein that is important for nucleocytoplasmic and cytosolic trafficking of the HIV-1 genomic RNA. Both hnRNP A2's interaction with HIV-1 RNA and its expression levels influence the activities of Rev in mediating nucleocytoplasmic export of the HIV-1 genomic RNA. While the lack of Rev expression during HIV-1 gene expression results in nuclear retention of HIV-1 genomic RNA, we show here by fluorescence in situ hybridization and fractionation studies that the genomic RNA translocates to the cytoplasm when hnRNP A2/B1 are depleted from cells. Polyribosome analyses revealed that the genomic RNA was shunted into a cytoplasmic, dense polyribosomal fraction. This fraction contained several RNA-binding proteins involved in viral gene expression and RNA trafficking but did not contain the translation initiation factor, eIF4G1. Amino acid incorporation into nascent polypeptides in this fraction was also greatly reduced, demonstrating that this fraction contains mRNAs that are poorly translated. These results demonstrate that hnRNP A2/B1 expression plays roles in the nuclear retention of the HIV-1 genomic RNA in the absence of Rev and in the release of the genomic RNA from translationally inactive, cytoplasmic RNP complexes.
hnRNP A2 是一种细胞蛋白,对 HIV-1 基因组 RNA 的核质和胞质转运很重要。hnRNP A2 与 HIV-1 RNA 的相互作用及其表达水平均影响 Rev 介导 HIV-1 基因组 RNA 核质输出的活性。虽然 HIV-1 基因表达期间 Rev 表达缺失会导致 HIV-1 基因组 RNA 在核内滞留,但我们通过荧光原位杂交和分级研究表明,当细胞内 hnRNP A2/B1 耗尽时,基因组 RNA 会转移到细胞质中。多核糖体分析显示,基因组 RNA 被转移到细胞质中,形成密集的多核糖体部分。该部分包含参与病毒基因表达和 RNA 转运的几个 RNA 结合蛋白,但不包含翻译起始因子 eIF4G1。该部分新生多肽的氨基酸掺入也大大减少,表明该部分包含翻译效率低的 mRNA。这些结果表明,hnRNP A2/B1 的表达在 Rev 缺失的情况下发挥了 HIV-1 基因组 RNA 在核内滞留的作用,并且在从翻译失活的细胞质 RNP 复合物中释放基因组 RNA 方面发挥了作用。