DELETION ANALYSIS OF A COMPLEX PROMOTER FOR A DEVELOPMENTALLY REGULATED GENE FROM BACILLUS-SUBTILIS
DELETION ANALYSIS OF A COMPLEX PROMOTER FOR A DEVELOPMENTALLY REGULATED GENE FROM BACILLUS-SUBTILIS
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DOI：
10.1016/s0022-2836(83)80023-0
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发表时间：
1983-01-01
影响因子：
5.6
通讯作者：
LOSICK, R
中科院分区：
文献类型：
--
作者：
BANNER, CDB;MORAN, CP;LOSICK, R
SpoVG是来自孢子形成细菌B的发育调控基因。枯草杆菌。spoVG的转录起始区由2个重叠启动子组成，其RNA合成起点相距10个碱基对。这些起始点分别被含有不同种类的B的2种形式的RNA聚合酶全酶利用。枯草σ因子构建了一系列从下游和上游方向延伸到spoVG启动子区的缺失突变。用这些突变的启动子进行的转录研究表明，spoVG启动子的功能边界从转录起始点的区域延伸到上游富含A + T的盒中，该盒位于下游起始位点之前的76-51个碱基对处。出乎意料地发现，spoVG启动子区在B中的高拷贝数质粒上的增殖。枯草芽孢杆菌通过在早期阶段损害发育而干扰孢子形成过程。这不是启动子扩增的一般效果，因为在2个其他强芽孢杆菌启动子的质粒上的繁殖对孢子形成几乎没有影响或没有影响。缺失分析表明，spoVG的区域引起孢子形成抑制密切相关的DNA序列所需的有效的启动子在体外利用。有人提出，spoVG的扩增是一种孢子形成特异性的调节蛋白，在转录起始区域或附近结合。
SpoVG is a developmentally regulated gene from the spore-forming bacterium B. subtilis. The transcription initiation region for spoVG consists of 2 overlapping promoters whose startpoints of RNA synthesis are 10 base pairs apart. These starpoints are separately utilized by 2 forms of RNA polymerase holoenzyme containing different species of B. subtilis .sigma. factor. A series of deletion mutations that extend into the spoVG promoter region from the downstream and from the upstream directions were constructed. Transcription studies with these mutated promoters showed that the functional boundaries of the spoVG promoters extended from the region of the transcription startpoints into an upstream A + T-rich box, which was located 76-51 base pairs preceding the downstream startsite. It was unexpectedly discovered that propagation of the spoVG promoter region on a high copy number plasmid in B. subtilis interferes with the process of sporulation by impairing development at an early stage. This was not a general effect of promoter amplification, since the propagation on plasmids of 2 other strong Bacillus promoters had little or no effect on spore formation. Deletion analysis established that the region of spoVG causing sporulation inhibition closely correlated with DNA sequences required for efficient promoter utilization in vitro. It is proposed that amplification of spoVG titrates a sporulation-specific regulatory protein that binds at or near the region of transcription initiation.