Expression and characterization of the trans-activator of HTLV-III/LAV virus.
Expression and characterization of the trans-activator of HTLV-III/LAV virus.
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DOI：
10.1126/science.3490693
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发表时间：
1986-11
期刊：
影响因子：
56.9
通讯作者：
C. M. Wright;B. Felber;H. Paskalis;G. Pavlakis
中科院分区：
文献类型：
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作者：
C. M. Wright;B. Felber;H. Paskalis;G. Pavlakis
人类嗜T淋巴细胞逆转录病毒HTLV-III/LAV编码一种增加病毒基因表达的反式激活因子。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征。假定的反式激活蛋白免疫沉淀过量生产稳定的细胞系，并显示迁移作为一个14千道尔顿的多肽十二烷基硫酸钠-聚丙烯酰胺凝胶。S1核酸酶作图实验表明，反式激活因子增加了从病毒长末端重复启动子转录的稳态信使RNA的水平。HTLV-III/LAV长末端重复的R区内的序列对于反式激活是必需的。信使RNA和蛋白质的定量显示，在CV1和HeLa细胞中，蛋白质的增加大于信使RNA的增加，表明不止一种机制负责反式激活，并且细胞类型特异性因素可能决定反式激活的最终水平。
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.