GGDEF-EAL domain proteins in bacterial physiology
GGDEF and EAL domain proteins were identified in the chromosome of many bacteria. GGDEF domain proteins are generally diguanyl cyclases which synthesize the secondary messenger cyclic di-GMP. EAL domain proteins are generally c-di-GMP specific phosphodiesterase which degrade c-di-GMP. C-di-GMP is involved in the regulation of multiple phenotypes in bacteria like biofilm formation, motility and virulence. In this work, we found that c-di-GMP inversely regulates biofilm formation and motility in bacteria such as Salmonella enterica serovar Typhimurium, Escherichia coli and Pseudomonas aeruginosa. Thereby, GGDEF domain proteins up-regulate c-di-GMP concentrations and the biofilm phenotype, while down regulating the motility phenotype. EAL domain proteins have the opposite effect. Consequently, cyclic di-GMP controls the transition from sessility to motility.
Phenotypic convergence is also observed when GGDEF domain proteins are exchanged between different species. Mutual complementation of GGDEF domain proteins mutants of S. Typhimurium and Yersinia pestis restored the respective phenotypes, cellulose biosynthesis and biofilm formation. In S. Typhimurium, the rdar morphotype, a multcellular behaviour that involves biofilm formation is regulated by c-di-GMP. The rdar morphotype is characterized by the expression of extracellular matrix components cellulose and curli fimbriae. CsgD is a major transcriptional regulator required for the activation of biosynthesis of cellulose and curli fimbriae. Systematic analysis of the 5 GGDEF, 7 EAL and 7 GGDEF-EAL domain proteins encoded on the S. Typhimurium chromosome indicates that these proteins do have overlapping, but never a redundant function. Mutant analysis revealed that the GGDEF-EAL domain proteins STM2123 and STM3388 enhance the expression of the transcriptional regulator CsgD at the transcriptional and post-transcriptional level and, consequently, rdar morphotype development, although no change in intracellular c-di-GMP levels is observed.
Chromosomal inactivation of AdrA does not have any significant effect on CsgD expression, although AdrA contributes to 60% of the basal level of c-di-GMP at a specific time point. On the other hand, four EAL or GGDEF-EAL domain proteins down-regulate the CsgD concentrations in the cell. While the c-di-GMP degraded by STM1703 is highly dedicated to regulate CsgD expression, c-di-GMP degraded by STM4264, STM3611 and STM1827 is only partially used to down-regulate CsgD expression. C-di-GMP metabolism is connected with other pathways in the cell like degradation of mRNAs. The expression of the EAL domain protein STM4264 dedicated to regulate CsgD expression and STM3611 dedicated to regulate motility are inversely regulated by exoribonuclease poly-nucleotide phosphorylase (PNPase), so that CsgD expression and motility are down-regulated. This finding explains the coherent regulation of opposing phenotypes by PNPase.
List of scientific papers
I. Simm R, Morr M, Kader A, Nimtz M, Römling U (2004). GGDEF and EAL domains inversely regulate cyclic di-GMP levels and transition from sessility to motility. Mol Microbiol. 53(4): 1123-34.
https://pubmed.ncbi.nlm.nih.gov/15306016
II. Simm R, Fetherston JD, Kader A, Römling U, Perry RD (2005). Phenotypic convergence mediated by GGDEF-domain-containing proteins. J Bacteriol. 187(19): 6816-23.
https://pubmed.ncbi.nlm.nih.gov/16166544
III. Kader A, Simm R, Gerstel U, Morr M, Römling U (2006). Hierarchical involvement of various GGDEF domain proteins in rdar morphotype development of Salmonella enterica serovar Typhimurium. Mol Microbiol. 60(3): 602-16.
https://pubmed.ncbi.nlm.nih.gov/16629664
IV. Simm R, Lusch A, Kader A, Andersson M, Römling U (2007). Role of EAL-containing proteins in multicellular behavior of Salmonella enterica serovar Typhimurium. J Bacteriol. 189(9): 3613-23. Epub 2007 Feb 23
https://pubmed.ncbi.nlm.nih.gov/17322315
V. Kader A, Römling U, Rhen M (2008). Polynucleotide phosphorylase promotes biofiolm formation and motility through differential regulation of EAL domain proteins in Salmonella enterica serovar Typhimurium. [Manuscript]
History
Defence date
2008-04-08Department
- Department of Microbiology, Tumor and Cell Biology
Publication year
2008Thesis type
- Doctoral thesis
ISBN
978-91-7357-565-2Number of supporting papers
5Language
- eng