Wright C M, Felber B K, Paskalis H, Pavlakis G N
Science. 1986 Nov 21;234(4779):988-92. doi: 10.1126/science.3490693.
The human T-lymphotropic retrovirus HTLV-III/LAV encodes a trans-activator that increases viral gene expression. We expressed this trans-activator in animal cells and studied its structural and functional characteristics. The putative trans-activator protein was immunoprecipitated from overproducing stable cell lines and shown to migrate as a 14-kilodalton polypeptide on sodium dodecyl sulfate-polyacrylamide gels. S1 nuclease mapping experiments showed that the trans-activator increases the levels of steady-state messenger RNA transcribed from the viral long terminal repeat promoter. Sequences within the R region of the HTLV-III/LAV long terminal repeat are essential for trans-activation. Quantitations of messenger RNA and protein showed that the protein increase was greater than the messenger RNA increase in CV1 and HeLa cells, indicating that more than one mechanism was responsible for the trans-activation and that cell type-specific factors may determine the final level of trans-activation.
人类嗜T淋巴细胞逆转录病毒HTLV-III/LAV编码一种反式激活因子,可增加病毒基因表达。我们在动物细胞中表达了这种反式激活因子,并研究了其结构和功能特性。从过量表达的稳定细胞系中免疫沉淀出推测的反式激活蛋白,在十二烷基硫酸钠-聚丙烯酰胺凝胶上显示其迁移为一条14千道尔顿的多肽。S1核酸酶图谱实验表明,反式激活因子可增加从病毒长末端重复启动子转录的稳态信使RNA水平。HTLV-III/LAV长末端重复序列R区域内的序列对于反式激活至关重要。信使RNA和蛋白质的定量分析表明,在CV1和HeLa细胞中,蛋白质增加量大于信使RNA增加量,这表明不止一种机制参与反式激活,且细胞类型特异性因子可能决定反式激活的最终水平。