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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Transcriptional inactivation of a regulatory site for replication of Vibrio cholerae chromosome II
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Transcriptional inactivation of a regulatory site for replication of Vibrio cholerae chromosome II

机译：转录灭活的霍乱弧菌染色体II复制的监管网站。

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摘要

The bacterium Vibrio cholerae has two chromosomes. The origin of replication of chromosome I is similar to that of Escherichia coli. The origin-containing region of chromosome II (oriCII) resembles replicons of plasmids such as P1, except for the presence of an additional gene, rctA [Egan, E. S. & Waldor, M. K. (2003) Cell 114, 521-530]. The oriCII region that includes the initiator gene, rctB, can function as a plasmid in E. coli. Here we show that RctB suffices for the oriCII-based plasmid replication, and rctA in cis or trans reduces the plasmid copy number, thereby serving as a negative regulator. The inhibitory activity could be overcome by increasing the concentration of RctB, suggesting that rctA titrates the initiator. Purified RctB bound to a DNA fragment carrying rctA, confirming that the two can interact. Although rctA apparently works as a titrating site, it is nonetheless transcribed. We find that the transcription attenuates the inhibitory activity of the gene, presumably by interfering with RctB binding. RctB, in turn, repressed the rctA promoter and, thereby, could control its own titration by modulating the transcription of rctA. This control circuit appears to be a putative novel mechanism for homeostasis of initiator availability.
机译：霍乱弧菌有两条染色体。染色体I的复制起点与大肠杆菌相似。染色体II的含起点区域（oriCII）类似于质粒P1的复制子，除了存在另外的基因rctA [Egan，E. S.＆Waldor，M. K.（2003）Cell 114，521-530]。包含启动子基因rctB的oriCII区可以在大肠杆菌中用作质粒。在这里，我们显示RctB足以满足基于oriCII的质粒复制，而rctA顺式或反式可降低质粒的拷贝数，从而充当负调节剂。抑制活性可以通过增加RctB的浓度来克服，这表明rctA滴定了引发剂。纯化的RctB与携带rctA的DNA片段结合，证实两者可以相互作用。尽管rctA显然可以作为滴定位点，但仍可以转录。我们发现，转录减弱了基因的抑制活性，大概是通过干扰RctB结合。反过来，RctB抑制了rctA启动子，从而可以通过调节rctA的转录来控制自身的滴定。该控制电路似乎是引发剂可用性动态平衡的推定新机制。

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