Slater S C, Lifsics M R, O'Donnell M, Maurer R
Department of Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106-4960.
J Bacteriol. 1994 Feb;176(3):815-21. doi: 10.1128/jb.176.3.815-821.1994.
DNA polymerase III holoenzyme is a multiprotein complex responsible for the bulk of chromosomal replication in Escherichia coli and Salmonella typhimurium. The catalytic core of the holoenzyme is an alpha epsilon theta heterotrimer that incorporates both a polymerase subunit (alpha; dnaE) and a proofreading subunit (epsilon; dnaQ). The role of theta is unknown. Here, we describe a null mutation of holE, the gene for theta. A strain carrying this mutation was fully viable and displayed no mutant phenotype. In contrast, a dnaQ null mutant exhibited poor growth, chronic SOS induction, and an elevated spontaneous mutation rate, like dnaQ null mutants of S. typhimurium described previously. The poor growth was suppressible by a mutation affecting alpha which was identical to a suppressor mutation identified in S. typhimurium. A double mutant null for both holE and dnaQ was indistinguishable from the dnaQ single mutant. These results show that the theta subunit is dispensable in both dnaQ+ and mutant dnaQ backgrounds, and that the phenotype of epsilon mutants cannot be explained on the basis of interference with theta function.
DNA聚合酶III全酶是一种多蛋白复合体,负责大肠杆菌和鼠伤寒沙门氏菌中的大部分染色体复制。全酶的催化核心是一个αεθ异源三聚体,它包含一个聚合酶亚基(α;dnaE)和一个校对亚基(ε;dnaQ)。θ的作用尚不清楚。在这里,我们描述了编码θ的基因holE的一个无效突变。携带这种突变的菌株完全可以存活,并且没有表现出突变表型。相比之下,一个dnaQ无效突变体生长不良、持续诱导SOS反应,并且自发突变率升高,就像之前描述的鼠伤寒沙门氏菌的dnaQ无效突变体一样。影响α的一个突变可以抑制生长不良,该突变与在鼠伤寒沙门氏菌中鉴定出的一个抑制突变相同。holE和dnaQ双突变体与dnaQ单突变体没有区别。这些结果表明,在dnaQ+和突变的dnaQ背景中,θ亚基都是可有可无的,并且ε突变体的表型不能基于对θ功能的干扰来解释。