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枯草芽孢杆菌中负责发育转录因子σK的基因是通过切除一个含有芽孢形成重组酶基因的可去除DNA元件而产生的。

The Bacillus subtilis gene for the development transcription factor sigma K is generated by excision of a dispensable DNA element containing a sporulation recombinase gene.

作者信息

Kunkel B, Losick R, Stragier P

机构信息

Biological Laboratories, Harvard University, Cambridge, Massachusetts 02138.

出版信息

Genes Dev. 1990 Apr;4(4):525-35. doi: 10.1101/gad.4.4.525.

Abstract

The structural gene (sigK) for the mother-cell RNA polymerase sigma-factor sigma K in Bacillus subtilis is a composite of two truncated genes, named spoIVCB and spoIIIC, which are brought together by site-specific recombination during sporulation. We now show that the recombination event is compartmentalized in that the mother cell, but not the forespore chromosome, undergoes rearrangement. We also show that spoIIIC (encoding the carboxy-terminal portion of sigma K) lies approximately 42 kb downstream of spoIVCB (encoding the amino-terminal portion) and that the joining of the truncated coding sequences is a reciprocal recombination event in which intervening DNA is deleted from the chromosome as a circle. The rearrangement is governed by the product of a gene named spoIVCA located in the excised DNA, as demonstrated by the observations (1) that the product of spoIVCA, but not the product of any other stage-IV sporulation gene tested, is required for the rearrangement, and (2) that the presence of a cloned copy of the rearranged sigK gene in the chromosome bypasses the requirement for the spoIVCA gene product in sporulation. Because cells engineered to contain an intact copy of sigK sporulate normally, we conclude that the sigK rearrangement is not essential for the control of gene expression during sporulation, and we infer the existence of an additional mechanism for restricting sigma K-directed transcription to the mother-cell chamber of the sporangium. Finally, the construction of a strain deleted for the entire sigK intervening sequence shows that the 42-kb element contains no genes essential for viability.

摘要

枯草芽孢杆菌中母细胞RNA聚合酶σ因子σK的结构基因(sigK)由两个截短的基因组成,分别命名为spoIVCB和spoIIIC,它们在芽孢形成过程中通过位点特异性重组结合在一起。我们现在表明,重组事件是分区进行的,即母细胞而非前芽孢染色体发生重排。我们还表明,spoIIIC(编码σK的羧基末端部分)位于spoIVCB(编码氨基末端部分)下游约42 kb处,截短编码序列的连接是一个相互重组事件,其中间隔DNA作为一个环从染色体上删除。重排由位于切除DNA中的一个名为spoIVCA的基因产物控制,这一结论基于以下观察结果:(1)重排需要spoIVCA的产物,但不需要测试的任何其他IV期芽孢形成基因的产物;(2)染色体中重排的sigK基因的克隆拷贝的存在绕过了芽孢形成过程中对spoIVCA基因产物的需求。由于经工程改造含有完整sigK拷贝的细胞能够正常形成芽孢,我们得出结论,sigK重排在芽孢形成过程中对基因表达的控制并非必不可少,并且我们推断存在一种额外的机制,将σK指导的转录限制在孢子囊的母细胞腔室中。最后,构建一个缺失整个sigK间隔序列的菌株表明,这个42 kb的元件不包含对生存力至关重要的基因。

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