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天蓝色链霉菌A3(2)的两个受发育调控的启动子,类似于其他细菌中主要的与运动性相关的启动子类别。

Two developmentally controlled promoters of Streptomyces coelicolor A3(2) that resemble the major class of motility-related promoters in other bacteria.

作者信息

Tan H, Chater K F

机构信息

John Innes Institute, John Innes Centre, Norwich, United Kingdom.

出版信息

J Bacteriol. 1993 Feb;175(4):933-40. doi: 10.1128/jb.175.4.933-940.1993.

Abstract

Experiments were designed to allow isolation of Streptomyces coelicolor promoters that depend on the whiG sporulation gene, which encodes a putative sigma factor important in the sporulation of aerial hyphae. The strategy, based on earlier evidence that sigma WhiG is limiting for sporulation (K. F. Chater, C. J. Burton, K. A. Plaskitt, M. J. Buttner, C. Méndez, and J. Helmann, Cell 59:133-143, 1989) was to seek DNA fragments that inhibit sporulation in aerial hyphae when present at a high copy number. In a suitable Sau3AI-generated library of DNA from S. coelicolor A3(2), two inserts were found to inhibit sporulation. Both inserts caused expression of the adjacent xylE reporter gene present in the vector in a developmentally normal strain of S. coelicolor, but there was no xylE expression in an otherwise isogenic whiG mutant. S1 nuclease protection experiments were done with RNAs isolated from these plasmid-bearing strains or from the wild-type strain lacking either recombinant plasmid. In each case, an apparent transcription start site was found upstream of an apparent open reading frame (ORF) and just downstream of sequences that resemble consensus features of promoters for motility-related genes in Bacillus subtilis and coliform bacteria. Such promoters depend on sigma factors (sigma D and sigma F, respectively) particularly similar to the deduced whiG gene product. Each of the putative whiG-dependent promoters is within an ORF that is upstream of, and potentially translationally coupled to, the putative whiG-dependent ORF (although use of one of the promoters would necessitate the use of a different start codon, further downstream). Thus, in unknown circumstances, the whiG-dependent ORFs may be expressed from a more remote promoter as part of a complex transcription unit.

摘要

实验旨在分离出天蓝色链霉菌中依赖于whiG孢子形成基因的启动子,该基因编码一种推测在气生菌丝孢子形成中起重要作用的σ因子。基于早期证据,即σWhiG对孢子形成具有限制作用(K. F. Chater、C. J. Burton、K. A. Plaskitt、M. J. Buttner、C. Méndez和J. Helmann,《细胞》59:133 - 143,1989),该策略是寻找当以高拷贝数存在时能抑制气生菌丝孢子形成的DNA片段。在一个合适的由天蓝色链霉菌A3(2)的Sau3AI酶切产生的DNA文库中,发现两个插入片段能抑制孢子形成。这两个插入片段都能使载体中相邻的xylE报告基因在天蓝色链霉菌发育正常的菌株中表达,但在其他方面同基因的whiG突变体中没有xylE表达。对从这些携带质粒的菌株或缺乏重组质粒的野生型菌株中分离得到的RNA进行了S1核酸酶保护实验。在每种情况下,都在一个明显的开放阅读框(ORF)上游、类似于枯草芽孢杆菌和大肠菌群细菌中与运动相关基因启动子的共有特征序列的下游发现了一个明显的转录起始位点。这样的启动子分别依赖于与推测的whiG基因产物特别相似的σ因子(分别为σD和σF)。每个推测的依赖于whiG的启动子都位于一个ORF内,该ORF在推测的依赖于whiG的ORF上游,并且可能在翻译上与之偶联(尽管使用其中一个启动子将需要使用更下游的不同起始密码子)。因此,在未知情况下,依赖于whiG的ORF可能作为一个复杂转录单元的一部分从更远处的启动子表达。

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