Huisman O, D'Ari R, Gottesman S
Proc Natl Acad Sci U S A. 1984 Jul;81(14):4490-4. doi: 10.1073/pnas.81.14.4490.
Blocks in DNA replication cause a rapid arrest of cell division in Escherichia coli. We have previously established that the function SfiA (SulA), induced under these conditions as part of the SOS response, is involved in this inhibition of division. To separate the effects of SfiA from those of other SOS functions, we have constructed a plac-sfiA operon fusion, permitting specific induction of SfiA protein by addition of the lac operon inducer isopropyl beta-D-thiogalactopyranoside (IPTG). In lon mutants, in which the unstable SfiA protein has a longer half-life, IPTG caused a rapid arrest of cell division. Under these conditions, there is no concomitant induction of the SOS response. IPTG also caused a rapid arrest of cell division in lon+ strains. These results demonstrate that induction of the SfiA protein is sufficient to cause inhibition of division. Mutations in the sfiB gene can suppress IPTG-induced SfiA-mediated inhibition of division. At higher SfiA concentrations, however, even sfiB mutants cease division; an additional mutation genetically inseparable from sfiB restores normal division. These observations reinforce the hypothesis that the SfiB protein, probably required for cell septation, is the target of action of the SfiA division inhibitor.
DNA复制受阻会导致大肠杆菌细胞分裂迅速停止。我们之前已经确定,在这些条件下作为SOS应答一部分而被诱导的SfiA(SulA)功能参与了这种对分裂的抑制作用。为了将SfiA的作用与其他SOS功能的作用区分开来,我们构建了一个plac-sfiA操纵子融合体,通过添加lac操纵子诱导剂异丙基-β-D-硫代半乳糖苷(IPTG)来特异性诱导SfiA蛋白。在lon突变体中,不稳定的SfiA蛋白具有更长的半衰期,IPTG导致细胞分裂迅速停止。在这些条件下,没有伴随SOS应答的诱导。IPTG在lon+菌株中也导致细胞分裂迅速停止。这些结果表明,SfiA蛋白的诱导足以导致对分裂的抑制。sfiB基因中的突变可以抑制IPTG诱导的SfiA介导的分裂抑制。然而,在较高的SfiA浓度下,即使是sfiB突变体也会停止分裂;一个与sfiB基因紧密连锁的额外突变可恢复正常分裂。这些观察结果强化了这样一种假说,即细胞分隔可能所需的SfiB蛋白是SfiA分裂抑制剂的作用靶点。