Peterlin B M, Luciw P A, Barr P J, Walker M D
Proc Natl Acad Sci U S A. 1986 Dec;83(24):9734-8. doi: 10.1073/pnas.83.24.9734.
The genome of human immunodeficiency virus encodes a protein that dramatically elevates amounts of viral proteins. The precise mechanism of this trans-activation remains to be established. It has been reported that trans-activation can occur without major changes in the levels of mRNA. We constructed recombinant plasmids containing those viral sequences required in cis for trans-activation linked to the chloramphenicol acetyltransferase gene. These plasmids were introduced into cultured cells in either the presence or absence of a second plasmid that directed expression of the viral trans-activator protein. Expression of the chloramphenicol acetyltransferase gene was measured at the level of protein (by enzymatic assay) and RNA (by ribonuclease protection and primer extension). Our results demonstrate that trans-activation is accompanied by large increases in mRNA levels; these increases may be sufficient to explain the elevated levels of trans-activated protein.
人类免疫缺陷病毒的基因组编码一种能显著提高病毒蛋白数量的蛋白质。这种反式激活的确切机制仍有待确定。据报道，反式激活可以在mRNA水平没有重大变化的情况下发生。我们构建了重组质粒，其中包含反式激活所需的顺式作用病毒序列，并与氯霉素乙酰转移酶基因相连。这些质粒在有或没有指导病毒反式激活蛋白表达的第二种质粒的情况下被导入培养细胞。通过酶促测定在蛋白质水平上测量氯霉素乙酰转移酶基因的表达，并通过核糖核酸酶保护和引物延伸在RNA水平上进行测量。我们的结果表明，反式激活伴随着mRNA水平的大幅增加；这些增加可能足以解释反式激活蛋白水平的升高。
