在CcpA和CodY中,它们调节了Streptococcus mutans的CRISPR-Cas系统
Da-Young Kang1, Andy Kim2, Jeong Nam Kim1,3
1Department of Integrated Biological Science, College of Natural Sciences, Pusan National University, Busan, Republic of Korea.
Microbiology spectrum
|June 27, 2023
概括
这项研究揭示了CcpA和CodY这两个关键调节器是如何控制Streptococcus mutans中的细菌CRISPR-Cas免疫系统的. 这种调节会影响果糖代谢和应激反应,这对于在不断变化的环境中生存至关重要.
科学领域:
- 微生物学 微生物学
- 细菌遗传学 细菌遗传学
- 分子生物学分子生物学
背景情况:
- 聚类定期间隔的短平行列重复 (CRISPR) 和与CRISPR相关的 (Cas) 基因在细菌中提供了对外来遗传元素的适应性免疫力.
- 口腔病原体Streptococcus mutans具有两个CRISPR-Cas位点,CRISPR1-Cas和CRISPR2-Cas,其转录调节仍然不完全理解.
- 全球调节者CcpA和CodY已知会影响碳水化合物和严格反应代谢.
研究的目的:
- 调查全球监管机构CcpA和CodY对Streptococcus mutans的转录调节的CRISPR-Cas操作.
- 阐明CcpA和CodY在调节CRISPR-Cas表达中的作用,以响应环境线索.
- 了解CRISPR-Cas系统活动对果糖代谢和严格反应的影响.
主要方法:
- 对CRISPR-Cas loci. 的促进区域和调节器结合点的计算预测.
- 电泳运动转移测定 (EMSA) 和DNase I足迹测定,以证实CcpA和CodY的结合.
- 构建和分析突变菌株 (例如, ΔccpA, ΔCR1cas, ΔCRDcas) 以评估各种条件下 (富含果糖,氧化应激,低pH值,mupirocin治疗) 的促进剂活性,果糖吸收和 (p) pGpp水平.
主要成果:
- CcpA直接与CRISPR-Cas loci的促进子区域结合,而CodY则表现出性相互作用.
- 果糖增强了CRISPR1-Cas促进剂活性,在富含果糖的条件下,在ΔccpA突变体中降低了CRISPR2-Cas活性.
- 删除CRISPR系统会降低果糖的吸收;CRISPR-Cas表达会影响严格反应期间的 (p) pGpp积累,并受到氧化,膜和低pH条件等环境压力的调节.
结论:
- 突变性链球菌的转录CRISPR-Cas系统是由CcpA和CodY的结合直接调节的.
- 这一规则对于协调CRISPR介导的免疫与碳水化合物代谢和能量消耗至关重要.
- 通过CcpA和CodY调控CRISPR-Cas,可以适应营养的可用性和环境压力,从而影响严格的反应和宿主生存.
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