Adachi T, Yamagata H, Tsukagoshi N, Udaka S
Department of Food Science and Technology, Faculty of Agriculture, Nagoya University, Japan.
J Bacteriol. 1989 Feb;171(2):1010-6. doi: 10.1128/jb.171.2.1010-1016.1989.
The genes encoding the major cell wall proteins, middle wall protein and outer wall protein, of Bacillus brevis 47 constitute a cotranscriptional unit (cwp [cell wall protein gene] operon). Primer extension assay of cwp operon transcripts showed the existence of six different 5' ends. This confirmed the results of the previous S1 nuclease protection assay and suggested the existence of several tandemly arranged promoters in the 5' region of the cwp operon. Promoter probe vectors carrying the Bacillus licheniformis alpha-amylase gene were constructed and used for deletion analysis of the 5' region. Three (P1, P2, and P3) of the six suggested promoters were shown to be located within three distinct fragments derived from the 5' region. The -35 and -10 regions of the P1 and P3 promoters resemble the consensus sequence recognized by the sigma-43-type RNA polymerase of Bacillus subtilis. The P2 promoter resembles only the consensus sequence in the -10 region. The P1 and P3 promoters were used to the same extents in Bacillus subtilis as in B. brevis, whereas the P2 promoter was used much less frequently in B. subtilis than in B. brevis. The P2 promoter is used constitutively in B. brevis 47 at all stages of growth, whereas P3 is used only at the exponential phase of growth. P2 could be a promoter of an unknown type that is preferentially used in B. brevis and might be responsible for the constitutive synthesis and secretion of the cell wall proteins into the medium at the stationary phase of growth.
短短芽孢杆菌47编码主要细胞壁蛋白、中层壁蛋白和外层壁蛋白的基因构成一个共转录单位（cwp[细胞壁蛋白基因]操纵子）。对cwp操纵子转录本进行的引物延伸分析显示存在6个不同的5'末端。这证实了先前S1核酸酶保护分析的结果，并表明在cwp操纵子的5'区域存在几个串联排列的启动子。构建了携带地衣芽孢杆菌α-淀粉酶基因的启动子探针载体，并用于对5'区域进行缺失分析。所推测的6个启动子中的3个（P1、P2和P3）位于源自5'区域的3个不同片段内。P1和P3启动子的-35区和-10区类似于枯草芽孢杆菌σ-43型RNA聚合酶识别的共有序列。P2启动子仅在-10区类似于共有序列。P1和P3启动子在枯草芽孢杆菌中的使用程度与在短短芽孢杆菌中的相同，而P2启动子在枯草芽孢杆菌中的使用频率远低于在短短芽孢杆菌中的使用频率。P2启动子在短短芽孢杆菌47生长的所有阶段均持续使用，而P3仅在生长的指数期使用。P2可能是一种未知类型的启动子，在短短芽孢杆菌中优先使用，可能负责在生长稳定期将细胞壁蛋白组成型合成并分泌到培养基中。
