1887

Abstract

To identify physiological processes affected by cAMP in the plant-symbiotic nitrogen-fixing α-proteobacterium Rm2011, cAMP levels were artificially increased by overexpression of its cognate adenylate/guanylate cyclase gene . This resulted in high accumulation of cAMP in the culture supernatant, decreased swimming motility and increased production of succinoglycan, an exopolysaccharide involved in host invasion. Weaker, similar phenotypic changes were induced by overexpression of and . Effects on swimming motility and succinoglycan production were partially dependent on encoding a cyclic AMP receptor-like protein. Transcriptome profiling of an -overexpressing strain identified 72 upregulated and 82 downregulated genes. A considerable number of upregulated genes are related to polysaccharide biosynthesis and osmotic stress response. These included succinoglycan biosynthesis genes, genes of the putative polysaccharide synthesis cluster and , the first gene of the operon encoding the FeuNPQ regulatory system. Downregulated genes were mostly related to respiration, central metabolism and swimming motility. Promoter-probe studies in the presence of externally added cAMP revealed 18 novel Clr-cAMP-regulated genes. Moreover, the addition of cGMP into the growth medium also promoted -dependent gene regulation. binding of Clr-cAMP and Clr-cGMP to the promoter regions of , and required the DNA motif (A/C/T)GT(T/C)(T/C/A)C (N) G(G/A)(T/A)ACA. Furthermore, and promoters were activated by Clr-cAMP/cGMP in as heterologous host. These findings suggest direct activation of these 7 genes by Clr-cAMP/cGMP.

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2016-10-01
2022-01-27
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