Yang X J, Hart C M, Grayhack E J, Roberts J W
Section of Biochemistry, Cornell University, Ithaca, New York 14853.
Genes Dev. 1987 May;1(3):217-26. doi: 10.1101/gad.1.3.217.
The gene Q protein of phage lambda is a transcription antiterminator that modifies RNA polymerase near the phage late gene promoter and thereby causes antitermination at distant sites. To define the site of action of Q protein, we have reconstructed the regulatory system on plasmids that allow the intracellular concentration of Q protein to be regulated, and that allow the effect of Q protein on transcription from variant promoter segments to be measured in vivo and in vitro. We show that DNA sequences essential for Q protein-mediated antitermination span the RNA start site, but do not extend beyond nucleotide 18 of the late RNA coding region. We also show that the modification that permits antitermination persists while RNA polymerase passes at least two terminators in vivo and in vitro.
噬菌体λ的基因Q蛋白是一种转录抗终止因子,它在噬菌体晚期基因启动子附近修饰RNA聚合酶,从而在远处位点导致抗终止。为了确定Q蛋白的作用位点,我们在质粒上重建了调控系统,该系统允许调节Q蛋白的细胞内浓度,并允许在体内和体外测量Q蛋白对来自变体启动子片段转录的影响。我们表明,Q蛋白介导的抗终止所必需的DNA序列跨越RNA起始位点,但不延伸到晚期RNA编码区的第18个核苷酸之外。我们还表明,允许抗终止的修饰在RNA聚合酶在体内和体外通过至少两个终止子时仍然存在。