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Unphosphorylated CsgD controls biofilm formation in Salmonella enterica serovar Typhimurium.
Mol Microbiol. 2010 Aug; 77(3):771-86.MM

Abstract

The transcriptional regulator CsgD of Salmonella enterica serovar Typhimurium (S. Typhimurium) is a major regulator of biofilm formation required for the expression of csgBA, which encodes curli fimbriae, and adrA, coding for a diguanylate cyclase. CsgD is a response regulator with an N-terminal receiver domain with a conserved aspartate (D59) as a putative target site for phosphorylation and a C-terminal LuxR-like helix-turn-helix DNA binding motif, but the mechanisms of target gene activation remained unclear. To study the DNA-binding properties of CsgD we used electrophoretic mobility shift assays and DNase I footprint analysis to show that unphosphorylated CsgD-His(6) binds specifically to the csgBA and adrA promoter regions. In vitro transcription analysis revealed that CsgD-His(6) is crucial for the expression of csgBA and adrA. CsgD-His(6) is phosphorylated by acetyl phosphate in vitro, which decreases its DNA-binding properties. The functional impact of D59 in vivo was demonstrated as S. Typhimurium strains expressing modified CsgD protein (D59E and D59N) were dramatically reduced in biofilm formation due to decreased protein stability and DNA-binding properties in the case of D59E. In summary, our findings suggest that the response regulator CsgD functions in its unphosphorylated form under the conditions of biofilm formation investigated in this study.

Authors+Show Affiliations

Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institutet, FE 280, 17177 Stockholm, Sweden.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20545866

Citation

Zakikhany, Katherina, et al. "Unphosphorylated CsgD Controls Biofilm Formation in Salmonella Enterica Serovar Typhimurium." Molecular Microbiology, vol. 77, no. 3, 2010, pp. 771-86.
Zakikhany K, Harrington CR, Nimtz M, et al. Unphosphorylated CsgD controls biofilm formation in Salmonella enterica serovar Typhimurium. Mol Microbiol. 2010;77(3):771-86.
Zakikhany, K., Harrington, C. R., Nimtz, M., Hinton, J. C., & Römling, U. (2010). Unphosphorylated CsgD controls biofilm formation in Salmonella enterica serovar Typhimurium. Molecular Microbiology, 77(3), 771-86. https://doi.org/10.1111/j.1365-2958.2010.07247.x
Zakikhany K, et al. Unphosphorylated CsgD Controls Biofilm Formation in Salmonella Enterica Serovar Typhimurium. Mol Microbiol. 2010;77(3):771-86. PubMed PMID: 20545866.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Unphosphorylated CsgD controls biofilm formation in Salmonella enterica serovar Typhimurium. AU - Zakikhany,Katherina, AU - Harrington,Carl R, AU - Nimtz,Manfred, AU - Hinton,Jay C D, AU - Römling,Ute, Y1 - 2010/06/09/ PY - 2010/6/16/entrez PY - 2010/6/16/pubmed PY - 2010/10/28/medline SP - 771 EP - 86 JF - Molecular microbiology JO - Mol Microbiol VL - 77 IS - 3 N2 - The transcriptional regulator CsgD of Salmonella enterica serovar Typhimurium (S. Typhimurium) is a major regulator of biofilm formation required for the expression of csgBA, which encodes curli fimbriae, and adrA, coding for a diguanylate cyclase. CsgD is a response regulator with an N-terminal receiver domain with a conserved aspartate (D59) as a putative target site for phosphorylation and a C-terminal LuxR-like helix-turn-helix DNA binding motif, but the mechanisms of target gene activation remained unclear. To study the DNA-binding properties of CsgD we used electrophoretic mobility shift assays and DNase I footprint analysis to show that unphosphorylated CsgD-His(6) binds specifically to the csgBA and adrA promoter regions. In vitro transcription analysis revealed that CsgD-His(6) is crucial for the expression of csgBA and adrA. CsgD-His(6) is phosphorylated by acetyl phosphate in vitro, which decreases its DNA-binding properties. The functional impact of D59 in vivo was demonstrated as S. Typhimurium strains expressing modified CsgD protein (D59E and D59N) were dramatically reduced in biofilm formation due to decreased protein stability and DNA-binding properties in the case of D59E. In summary, our findings suggest that the response regulator CsgD functions in its unphosphorylated form under the conditions of biofilm formation investigated in this study. SN - 1365-2958 UR - https://www.unboundmedicine.com/medline/citation/20545866/Unphosphorylated_CsgD_controls_biofilm_formation_in_Salmonella_enterica_serovar_Typhimurium_ L2 - https://doi.org/10.1111/j.1365-2958.2010.07247.x DB - PRIME DP - Unbound Medicine ER -