York K, Kenney T J, Satola S, Moran C P, Poth H, Youngman P
Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia 19104.
J Bacteriol. 1992 Apr;174(8):2648-58. doi: 10.1128/jb.174.8.2648-2658.1992.
The spoIIE operon is a developmentally regulated transcription unit activated in the second hour of sporulation in Bacillus subtilis. Its promoter has an unusual structure, containing sequences which conform perfectly to the consensus for vegetative promoters recognized by sigma A-associated RNA polymerase (E sigma A), but with a spacing of 21 bp between the apparent -10 and -35 elements instead of the 17- or 18-bp spacing typical of promoters utilized by E sigma A. Mutations introduced into the apparent -10 element affected transcription in a manner consistent with its functioning as a polymerase recognition sequence. The deleterious effect of one -10 mutation was also suppressed in an allele-specific manner by a mutation in sigA known to suppress analogous -10 mutations in conventional vegetative promoters recognized by E sigma A. Similar suppression experiments failed to provide evidence for a direct interaction between E sigma A and the "-35-like" element, however, and DNase I protection experiments suggested instead that the Spo0A protein binds to a site overlapping this -35-like hexamer. Moreover, the effects of mutations within the -35-like hexamer on the binding of Spo0A in vitro paralleled their effects on transcription in vivo. We suggest that spoIIE belongs to a class of early-intermediate sporulation genes whose transcription by E sigma A is activated by the Spo0A protein.
spoIIE操纵子是一个受发育调控的转录单元,在枯草芽孢杆菌芽孢形成的第二个小时被激活。其启动子具有不寻常的结构,包含与由σA相关RNA聚合酶(EσA)识别的营养型启动子的共有序列完全一致的序列,但在明显的-10和-35元件之间的间距为21 bp,而不是EσA使用的启动子典型的17或18 bp间距。引入到明显的-10元件中的突变以与其作为聚合酶识别序列的功能一致的方式影响转录。一个-10突变的有害效应也以等位基因特异性的方式被sigA中的一个突变所抑制,该突变已知可抑制EσA识别的传统营养型启动子中的类似-10突变。然而,类似的抑制实验未能提供EσA与“-35样”元件之间直接相互作用的证据,并且DNase I保护实验反而表明Spo0A蛋白结合到与这个-35样六聚体重叠的位点。此外,-35样六聚体内的突变对体外Spo0A结合的影响与其对体内转录的影响平行。我们认为spoIIE属于一类早期-中期芽孢形成基因,其由EσA进行的转录被Spo0A蛋白激活。