Ivey-Hoyle M, Rosenberg M
Department of Gene Expression Sciences, SmithKline Beecham Pharmaceuticals, King of Prussia, Pennsylvania 19406.
Mol Cell Biol. 1990 Dec;10(12):6152-9. doi: 10.1128/mcb.10.12.6152-6159.1990.
Expression of the human immunodeficiency virus (HIV) structural proteins in mammalian cells is regulated posttranscriptionally by the viral Rev protein. Rev has been shown to trans-activate expression by relieving the nuclear sequestration of RNAs containing viral gag or env coding regions. We have studied the effects of Rev on expression of the HIV type 1 env gene in Drosophila melanogaster cells. We demonstrated that synthesis of the gp160 envelope protein was fully Rev dependent; that is, gp160 was produced only when Rev function was coexpressed in the cell. Analysis of total cellular RNA indicated that Rev did not significantly affect the overall levels of gp160 RNA production. Instead, mRNA encoding gp160 was found in the cytoplasm only in cells expressing Rev, whereas in cells lacking Rev, this RNA was present only in the nucleus. Furthermore, comparison of these results with the previously demonstrated Rev-independent expression of gp120 envelope protein with this system indicated that information contained in the gp41 coding region appears to be critical to the selective nuclear retention of gp160 transcripts in the absence of Rev. Our results clearly demonstrate that the mechanism of Rev action is conserved in the insect cell system, and, thus, Rev must function via cellular machinery common to most, if not all, higher cell systems.
人类免疫缺陷病毒(HIV)结构蛋白在哺乳动物细胞中的表达受病毒Rev蛋白的转录后调控。Rev已被证明可通过解除对含有病毒gag或env编码区的RNA的核隔离来反式激活表达。我们研究了Rev对果蝇细胞中HIV - 1 env基因表达的影响。我们证明gp160包膜蛋白的合成完全依赖于Rev;也就是说,只有当Rev功能在细胞中共表达时才会产生gp160。对总细胞RNA的分析表明,Rev对gp160 RNA的总体产生水平没有显著影响。相反,编码gp160的mRNA仅在表达Rev的细胞的细胞质中被发现,而在缺乏Rev的细胞中,这种RNA仅存在于细胞核中。此外,将这些结果与先前证明的该系统中gp120包膜蛋白的Rev非依赖性表达进行比较表明,gp41编码区中包含的信息似乎对在没有Rev的情况下gp160转录本的选择性核滞留至关重要。我们的结果清楚地表明,Rev的作用机制在昆虫细胞系统中是保守的,因此,Rev必须通过大多数(如果不是全部)高等细胞系统共有的细胞机制发挥作用。