Favaro J P, Maldarelli F, Arrigo S J, Schmidt M G
Department of Microbiology and Immunology, Medical University of South Carolina, Charleston, South Carolina, 29425-2230, USA.
Virology. 1999 Mar 15;255(2):237-49. doi: 10.1006/viro.1998.9584.
Human immunodeficiency virus type 1 (HIV-1) proteins are expressed from both intron-containing and completely spliced RNAs. Rev, an HIV-1 regulatory protein, is necessary for the expression of intron-containing RNAs. The effect of Rev on the subcellular localization of intron-containing HIV-1 RNA was examined by in situ RNA hybridization. In the presence of Rev, intron-containing HIV-1 RNA accumulated at the nuclear membrane and within the cytoplasm of transfected cells. In the absence of Rev, intron-containing HIV-1 RNA accumulated within the nucleus. In approximately 20% of the cells transfected in the absence of Rev, intron-containing HIV-1 RNA was also found in the cytoplasm. Differences in the subcytoplasmic localization of intron-containing HIV-1 RNA in the presence and absence of Rev were not observed using in situ RNA hybridization. To determine the effect of Rev on RNA localization within the cytoplasm, an extensive fractionation protocol involving both hypotonic and detergent lysis was used. In the presence of Rev, 40.9 +/- 4.6% of the cytoplasmic intron-containing HIV-1 RNA was released by hypotonic lysis. A similar fractionation profile was seen for several other translated viral and cellular RNAs. However, in the absence of Rev, only 16.5 +/- 5.1% of the cytoplasmic intron-containing HIV-1 RNA was released on hypotonic lysis (P < 0. 005). Thus the cytoplasmic fractionation pattern of this RNA was altered in the absence of Rev.
1型人类免疫缺陷病毒（HIV-1）蛋白由含内含子的RNA和完全剪接的RNA表达。Rev是一种HIV-1调节蛋白，对于含内含子RNA的表达是必需的。通过原位RNA杂交检测了Rev对含内含子的HIV-1 RNA亚细胞定位的影响。在有Rev存在的情况下，含内含子的HIV-1 RNA在转染细胞的核膜和细胞质中积累。在没有Rev的情况下，含内含子的HIV-1 RNA在细胞核内积累。在没有Rev的情况下转染的细胞中，约20%的细胞在细胞质中也发现了含内含子的HIV-1 RNA。使用原位RNA杂交未观察到有或没有Rev时含内含子的HIV-1 RNA在细胞质亚定位上的差异。为了确定Rev对细胞质内RNA定位的影响，使用了一种涉及低渗和去污剂裂解的广泛分级分离方案。在有Rev存在的情况下，40.9±4.6%的细胞质中含内含子的HIV-1 RNA通过低渗裂解释放出来。其他几种翻译的病毒和细胞RNA也观察到了类似的分级分离情况。然而，在没有Rev的情况下，低渗裂解时只有16.5±5.1%的细胞质中含内含子的HIV-1 RNA被释放出来（P<0.005）。因此，在没有Rev的情况下，这种RNA的细胞质分级分离模式发生了改变。
