Nanamiya H, Fugono N, Asai K, Doi R H, Kawamura F
Laboratory of Molecular Genetics, College of Science, Rikkyo University, Tokyo, Japan.
FEMS Microbiol Lett. 2000 Nov 15;192(2):237-41. doi: 10.1111/j.1574-6968.2000.tb09388.x.
We isolated a temperature-sensitive sporulation defective mutant of the sigA gene, encoding a major sigma factor, sigma(A) protein, in Bacillus subtilis, and designated it as sigA21. The sigA21 mutation caused a single-amino acid substitution, E314K, in region 4 of the sigma(A) protein. In this mutant, expression of the spoIIG gene, whose transcription depends on both sigma(A) and the phosphorylated Spo0A protein, Spo0A approximately P, a major transcription factor during early stages of sporulation, was greatly reduced at 43 degrees C. To obtain further information on the mechanism of sigma(A) function during the early spore development, we isolated a spontaneous sporulation-proficient suppressor mutant at 43 degrees C. This extragenic suppressor mutation was mapped within the rpoB gene, encoding the beta subunit of RNA polymerase, and was found to have a single-amino acid substitution, A863G. In this mutant, the expression of the spoIIG is partially restored at 43 degrees C.
我们在枯草芽孢杆菌中分离出了编码主要σ因子σ(A)蛋白的sigA基因的温度敏感型孢子形成缺陷突变体,并将其命名为sigA21。sigA21突变在σ(A)蛋白的第4区域导致了单个氨基酸取代,即E314K。在这个突变体中,spoIIG基因的表达在43℃时大幅降低,spoIIG基因的转录依赖于σ(A)和磷酸化的Spo0A蛋白Spo0AP,Spo0AP是孢子形成早期的主要转录因子。为了获得关于σ(A)在早期孢子发育过程中功能机制的更多信息,我们分离出了一个在43℃时自发产生的孢子形成能力恢复的抑制突变体。这个基因外抑制突变定位在编码RNA聚合酶β亚基的rpoB基因内,并且发现有单个氨基酸取代,即A863G。在这个突变体中,spoIIG的表达在43℃时部分恢复。