Departamento de Biología Molecular, Área de Bioquímica y Biología Molecular, Universidad de León, Campus de Vegazana, 24071 León, Spain.
Biosci Rep. 2013 May 24;33(3):e00038. doi: 10.1042/BSR20130018.
We have shown previously that Escherichia coli K92 produces two different capsular polymers known as CA (colanic acid) and PA (polysialic acid) in a thermoregulated manner. The complex Rcs phosphorelay is largely related to the regulation of CA synthesis. Through deletion of rscA and rscB genes, we show that the Rcs system is involved in the regulation of both CA and PA synthesis in E. coli K92. Deletion of either rcsA or rcsB genes resulted in decreased expression of cps (CA biosynthesis cluster) at 19°C and 37°C, but only CA production was reduced at 19°C. Concerning PA, both deletions enhanced its synthesis at 37°C, which does not correlate with the reduced kps (PA biosynthesis cluster) expression observed in the rcsB mutant. Under this condition, expression of the nan operon responsible for PA catabolism was greatly reduced. Although RcsA and RcsB acted as negative regulators of PA synthesis at 37°C, their absence did not reestablish PA expression at low temperatures, despite the deletion of rcsB resulting in enhanced kps expression. Finally, our results revealed that RcsB controlled the expression of several genes (dsrA, rfaH, h-ns and slyA) involved in the thermoregulation of CA and PA synthesis, indicating that RcsB is part of a complex regulatory mechanism governing the surface appearance in E. coli.
我们之前已经表明,大肠杆菌 K92 以热调节的方式产生两种不同的荚膜聚合物,即 CA(黏多糖酸)和 PA(聚唾液酸)。复杂的 Rcs 磷酸传递系统与 CA 合成的调节密切相关。通过删除 rscA 和 rscB 基因,我们表明 Rcs 系统参与了大肠杆菌 K92 中 CA 和 PA 合成的调节。rcsA 或 rscB 基因的缺失导致 cps(CA 生物合成簇)在 19°C 和 37°C 时的表达降低,但仅在 19°C 时降低了 CA 的产生。关于 PA,两个缺失都增强了其在 37°C 时的合成,这与 rcsB 突变体中观察到的 kps(PA 生物合成簇)表达减少无关。在这种情况下,负责 PA 分解代谢的 nan 操纵子的表达大大降低。尽管 RcsA 和 RcsB 在 37°C 时作为 PA 合成的负调节剂,但它们的缺失并没有在低温下重新建立 PA 表达,尽管 rcsB 的缺失导致 kps 表达增强。最后,我们的结果表明,RcsB 控制着几个基因(dsrA、rfaH、h-ns 和 slyA)的表达,这些基因参与 CA 和 PA 合成的热调节,表明 RcsB 是控制大肠杆菌表面外观的复杂调节机制的一部分。