首页> 美国卫生研究院文献>Proceedings of the National Academy of Sciences of the United States of America >Stringent control and growth-rate-dependent control have nonidentical promoter sequence requirements.
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Stringent control and growth-rate-dependent control have nonidentical promoter sequence requirements.

机译:严格控制和生长速率依赖性控制具有不同的启动子序列要求。

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摘要

Escherichia coli uses at least two regulatory systems, stringent control and growth-rate-dependent control, to adjust rRNA output to amino acid availability and the steady-state growth rate, respectively. We examined transcription from rrnB P1 promoters containing or lacking the cis-acting UP element and FIS protein binding sites after amino acid starvation. The "core promoter" responds to amino acid starvation like the full-length wild-type promoter; thus, neither the UP element nor FIS plays a role in stringent control. To clarify the relationship between growth-rate-dependent regulation and stringent control, we measured transcription from growth-rate-independent promoters during amino acid starvation. Four rrnB P1 mutants defective for growth-rate control and two other growth-rate-independent promoters (rrnB P2 and pS10) still displayed stringent regulation. Thus, the two systems have different promoter determinants, consistent with the idea that they function by different mechanisms. Two mutations disrupted stringent control of rrnB P1: (i) a multiple base change in the "discriminator" region between the -10 hexamer and the transcription start site and (ii) a double substitution making the promoter resemble the E sigma 70 consensus promoter. These results have important implications for the mechanisms of both stringent control and growth-rate-dependent control of rRNA transcription.
机译:大肠杆菌至少使用两种调节系统,即严格控制和生长速率依赖性控制,以分别将rRNA输出调节为氨基酸可利用度和稳态生长率。我们检查了氨基酸饥饿后从rrnB P1启动子的转录,该启动子含有或缺乏顺式作用的UP元件和FIS蛋白结合位点。 “核心启动子”像全长野生型启动子一样响应氨基酸饥饿。因此,UP元素和FIS都不在严格控制中发挥作用。为了阐明生长速率依赖性调节与严格控制之间的关系,我们在氨基酸饥饿期间测量了生长速率依赖性启动子的转录。四个rrnB P1突变体对生长速率控制有缺陷,另外两个不依赖生长速率的启动子(rrnB P2和pS10)仍然显示严格的调控。因此,这两个系统具有不同的启动子决定簇,这与它们通过不同机制起作用的想法一致。两种突变破坏了对rrnB P1的严格控制:(i)-10六聚体和转录起始位点之间“鉴别​​子”区域的多碱基变化,以及(ii)使启动子类似于E sigma 70共有启动子的双取代。这些结果对rRNA转录的严格控制和生长速率依赖性控制的机制具有重要意义。

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