Pedersen L B, Ragkousi K, Cammett T J, Melly E, Sekowska A, Schopick E, Murray T, Setlow P
Department of Biochemistry, University of Connecticut Health Center, Farmington, CT 06032, USA.
Gene. 2000 Apr 4;246(1-2):187-96. doi: 10.1016/s0378-1119(00)00084-6.
The Bacillus subtilis genome sequencing project [Kunst et al., Nature 390 (1997) 249-256] identified ywhE as a gene that potentially encodes a high-molecular-weight class A penicillin-binding protein. Analysis of the expression of a translational ywhE-lacZ fusion showed that ywhE expression is sporulation-specific, and is controlled predominantly by the forespore-specific sigma factor sigma(F), and to a lesser extent by sigma(G). Primer extension analysis identified two transcription start sites located 26 and 27 nucleotides upstream of the ywhE translational initiation codon. Sequences located in the -10 and -35 regions relative to the transcription start sites showed good homology to the consensus sequences for promoter elements of sigma(F)-dependent genes. An insertional mutation in ywhE had no significant effect on growth, morphology, and sporulation, and ywhE spores had normal heat-resistance, cortex structure, and germination and outgrowth properties. However, overexpression of ywhE in Escherichia coli resulted in cell lysis.
枯草芽孢杆菌基因组测序项目[孔斯特等人，《自然》390(1997)249 - 256]将ywhE鉴定为一个可能编码高分子量A类青霉素结合蛋白的基因。对ywhE - lacZ翻译融合体表达的分析表明，ywhE的表达具有芽孢形成特异性，主要受前芽孢特异性σ因子σ(F)控制，在较小程度上受σ(G)控制。引物延伸分析确定了位于ywhE翻译起始密码子上游26和27个核苷酸处的两个转录起始位点。相对于转录起始位点，位于 - 10和 - 35区域的序列与σ(F)依赖性基因启动子元件的共有序列具有良好的同源性。ywhE中的插入突变对生长、形态和芽孢形成没有显著影响，ywhE芽孢具有正常的耐热性、皮层结构以及萌发和生长特性。然而，ywhE在大肠杆菌中的过表达导致细胞裂解。
