Bent A F, Signer E R
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
J Bacteriol. 1990 Jul;172(7):3559-68. doi: 10.1128/jb.172.7.3559-3568.1990.
sigma 32, the product of the Escherichia coli rpoH locus, is an alternative RNA polymerase sigma factor utilized to express heat shock genes upon a sudden rise in temperature. E. coli K165 [rpoH165(Am) supC(Ts)] is temperature sensitive for growth and does not induce heat shock protein synthesis. We have isolated a locus from Rhizobium meliloti called suhR that allows E. coli K165 to grow at high temperature and induce heat shock protein synthesis. R. meliloti suhR mutants were viable and symbiotically effective. suhR was found to have no DNA or derived amino acid sequence similarity to the genes of previously sequenced sigma factors or other data base entries, although a helix-turn-helix DNA-binding protein motif is present. suhR did not restore the phenotypic defects of delta rpoH E. coli; suppression of the E. coli K165 phenotype is thus likely to involve E. coli sigma 32. Western immunoblots showed that suhR caused an approximately twofold elevation of sigma 32 levels in K165; RNA blots indicated that rpoH mRNA level and stability were not altered. Stabilization of sigma 32 protein and increased rpoH mRNA translation are thus the most probable mechanisms of suppression.
σ32是大肠杆菌rpoH基因座的产物,是一种替代性RNA聚合酶σ因子,用于在温度突然升高时表达热休克基因。大肠杆菌K165 [rpoH165(Am) supC(Ts)] 对生长温度敏感,且不诱导热休克蛋白合成。我们从苜蓿根瘤菌中分离出一个名为suhR的基因座,它能使大肠杆菌K165在高温下生长并诱导热休克蛋白合成。苜蓿根瘤菌suhR突变体是存活的且具有共生效应。尽管存在一个螺旋-转角-螺旋DNA结合蛋白基序,但发现suhR与先前测序的σ因子基因或其他数据库条目没有DNA或推导的氨基酸序列相似性。suhR不能恢复缺失rpoH的大肠杆菌的表型缺陷;因此,对大肠杆菌K165表型的抑制可能涉及大肠杆菌σ32。蛋白质免疫印迹显示,suhR使K165中的σ32水平升高了约两倍;RNA印迹表明rpoH mRNA水平和稳定性没有改变。因此,σ32蛋白的稳定和rpoH mRNA翻译增加是最可能的抑制机制。