在大肠杆菌中运激活的fucAO的综合性表征
Zhongge Zhang1, Jialu Huo1, Juan Velo1
1Department of Molecular Biology, School of Biological Sciences, University of California at San Diego, 9500 Gilman Dr, La Jolla, CA 92093-0116, USA.
大肠杆菌中的fucAO操作子由糖激活,需要FucR和Crp-cAMP. FucR促进了Crp-cAMP与促进体的结合,使RNA聚合酶能够招募转录.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 基因规则 基因规则
背景情况:
- 野生型大肠杆菌无法代谢L-1,2-二醇.
- fucAO操作子是fucose regulon的一部分,在没有L-fucose的情况下通常是无声的.
- 关于fucAO操作子的转录调节的信息有限.
研究的目的:
- 为了研究 Escherichia coli 中 fucAO 操作子的转录调节.
- 确定促进区域和转录开始地点.
- 阐明FucR和Crp-cAMP在操作子激活中的作用.
主要方法:
- 证实了fucAO操作子的inducibility通过fucose.
- 5'RACE用于确定转录起点 (TSS).
- 分析FucR和Crp结合位点及其在转录中的作用.
主要成果:
- fucAO操作子是高度可诱导的fucose,主要是通过一个上游促进器.
- 确定了实际的TSS,与之前的假设有所不同.
- 运激活取决于FucR和Crp-cAMP,并确定了特定的结合部位.
- 在转录中,Crp-cAMP比FucR发挥更重要的作用.
结论:
- FucR促进了Crp-cAMP与上游站点的结合.
- 结合crp-cAMP对于招募RNA聚合酶和激活fucAO促进体至关重要.
- 这种机制解释了fucAO操作子对fucose的反应中的转录调节.
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