Viglianti G A, Sharma P L, Mullins J I
Department of Cancer Biology, Harvard University School of Public Health, Boston, Massachusetts 02115.
J Virol. 1990 Sep;64(9):4207-16. doi: 10.1128/JVI.64.9.4207-4216.1990.
The human and simian immunodeficiency viruses encode at least six gene products that apparently serve regulatory functions. To evaluate the regulation of simian immunodeficiency virus gene expression at the level of RNA splicing, we used the polymerase chain reaction to amplify and clone cDNAs corresponding to a large array of mRNAs from infected cells. We identified mRNAs that used splice acceptor sites upstream of the initiator codons for tat, rev, vpr, nef, vif, and vpx, suggesting that these proteins may be expressed from different mRNAs. We also provide hybridization data suggesting that the same splice acceptor site may be used for both rev and env mRNAs. Furthermore, we isolated both tat and rev cDNAs that utilized three alternative splice acceptor sites at the start of coding exon 2, indicating that different versions of these proteins may be encoded. Finally, approximately 10 to 20% of simian immunodeficiency virus mRNAs spliced an intron from their untranslated 5' ends, and sequences contained within this intron constituted a portion of the tat-responsive TAR element. Thus, alternative pre-mRNA splicing adds a level of complexity to simian immunodeficiency virus expression, which may affect several levels of gene regulation.
人类免疫缺陷病毒和猿猴免疫缺陷病毒编码至少六种明显具有调节功能的基因产物。为了在RNA剪接水平评估猿猴免疫缺陷病毒基因表达的调控,我们使用聚合酶链反应来扩增和克隆与来自感染细胞的大量mRNA相对应的cDNA。我们鉴定出了使用tat、rev、vpr、nef、vif和vpx起始密码子上游剪接受体位点的mRNA,这表明这些蛋白质可能由不同的mRNA表达。我们还提供了杂交数据,表明rev和env mRNA可能使用相同的剪接受体位点。此外,我们分离出了在编码外显子2起始处利用三个可变剪接受体位点的tat和rev cDNA,这表明这些蛋白质可能编码不同的版本。最后,大约10%至20%的猿猴免疫缺陷病毒mRNA从其未翻译的5'端剪接了一个内含子,并且该内含子中包含的序列构成了tat反应性TAR元件的一部分。因此,可变前体mRNA剪接增加了猿猴免疫缺陷病毒表达的复杂性,这可能会影响基因调控的多个层面。