Kumar Amrita, Brannigan James A, Moran Charles P
Department of Microbiology & Immunology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
J Bacteriol. 2004 Feb;186(4):1078-83. doi: 10.1128/JB.186.4.1078-1083.2004.
At the onset of endospore formation in Bacillus subtilis, the DNA binding protein Spo0A activates transcription from two types of promoters. The first type includes the spoIIG and spoIIE promoters, which are used by sigma(A)-RNA polymerase, whereas the second type includes the spoIIA promoter, which is used by RNA polymerase containing the secondary sigma factor sigma(H). Previous genetic analyses have identified specific amino acids in alpha-helix E of Spo0A that are important for activation of Spo0A-dependent, sigma(A)-dependent promoters. However, these amino acids are not required for activation of the sigma(H)-dependent spoIIA promoter. We now report the effects of additional single-amino-acid substitutions and the effects of deletions in alpha-helix E. The effects of alanine substitutions revealed one new position (239) in Spo0A that appears to be specifically required for activation of the sigma(A)-dependent promoters. Based on the effects of a deletion mutation, we suggest that alpha-helix E in Spo0A is not directly involved in interaction with sigma(H)-RNA polymerase.
在枯草芽孢杆菌芽孢形成开始时,DNA结合蛋白Spo0A激活两种类型启动子的转录。第一种类型包括spoIIG和spoIIE启动子,它们由σ(A)-RNA聚合酶使用,而第二种类型包括spoIIA启动子,它由含有二级σ因子σ(H)的RNA聚合酶使用。先前的遗传分析已经确定了Spo0A的α螺旋E中的特定氨基酸,这些氨基酸对于激活Spo0A依赖的、σ(A)依赖的启动子很重要。然而,这些氨基酸对于激活σ(H)依赖的spoIIA启动子不是必需的。我们现在报告α螺旋E中额外单氨基酸取代的影响以及缺失的影响。丙氨酸取代的影响揭示了Spo0A中一个新的位置(239),该位置似乎是激活σ(A)依赖启动子所特需的。基于缺失突变的影响,我们认为Spo0A中的α螺旋E不直接参与与σ(H)-RNA聚合酶的相互作用。