Transcription of the Alginate Operon in Pseudomonas aeruginosa Is Regulated by c-di-GMP
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Transcription of the Alginate Operon in Pseudomonas aeruginosa Is Regulated by c-di-GMP. / Liang, Ziwei; Rybtke, Morten; Kragh, Kasper Nørskov; Johnson, Owen; Schicketanz, Muriel; Zhang, Yong Everett; Andersen, Jens Bo; Tolker-Nielsen, Tim.
In: Microbiology Spectrum, Vol. 10, No. 4, e00675-22, 2022.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Transcription of the Alginate Operon in Pseudomonas aeruginosa Is Regulated by c-di-GMP
AU - Liang, Ziwei
AU - Rybtke, Morten
AU - Kragh, Kasper Nørskov
AU - Johnson, Owen
AU - Schicketanz, Muriel
AU - Zhang, Yong Everett
AU - Andersen, Jens Bo
AU - Tolker-Nielsen, Tim
PY - 2022
Y1 - 2022
N2 - Overproduction of the exopolysaccharide alginate contributes to the pathogenicity and antibiotic tolerance of Pseudomonas aeruginosa in chronic infections. The second messenger, c-di-GMP, is a positive regulator of the production of various biofilm matrix components and is known to regulate alginate synthesis at the posttranslational level in P. aeruginosa. We provide evidence that c-di-GMP also regulates transcription of the alginate operon in P. aeruginosa. Previous work has shown that transcription of the alginate operon is regulated by nine different proteins, AmrZ, AlgP, IHF alpha, IHF beta, CysB, Vfr, AlgR, AlgB, and AlgQ, and we investigated if some of these proteins function as a c-di-GMP effector. We found that deletion of algP, algQ, IHF alpha, and IHF beta had only a marginal effect on the transcription of the alginate operon. Deletion of vfr and cysB led to decreased transcription of the alginate operon, and the dependence of the c-di-GMP level was less pronounced, indicating that Vfr and CysB could be partially required for c-di-GMP-mediated regulation of alginate operon transcription. Our experiments indicated that the AmrZ, AlgR, and AlgB proteins are absolutely required for transcription of the alginate operon. However, differential radial capillary action of ligand assay (DRaCALA) and site-directed mutagenesis indicated that c-di-GMP does not bind to any of the AmrZ, AlgR, and AlgB proteins. IMPORTANCE The proliferation of alginate-overproducing P. aeruginosa variants in the lungs of cystic fibrosis patients often leads to chronic infection. The alginate functions as a biofilm matrix that protects the bacteria against host immune defenses and antibiotic treatment. Knowledge about the regulation of alginate synthesis is important in order to identify drug targets for the development of medicine against chronic P. aeruginosa infections. We provide evidence that c-di-GMP positively regulates transcription of the alginate operon in P. aeruginosa. Moreover, we revisited the role of the known alginate regulators, AmrZ, AlgP, IHF alpha, IHF beta, CysB, Vfr, AlgR, AlgB, and AlgQ, and found that their effect on transcription of the alginate operon is highly varied. Deletion of algP, algQ, IHF alpha, or IHF beta only had a marginal effect on transcription of the alginate operon, whereas deletion of vfr or cysB led to decreased transcription and deletion of amrZ, algR, or algB abrogated transcription.The proliferation of alginate-overproducing P. aeruginosa variants in the lungs of cystic fibrosis patients often leads to chronic infection. The alginate functions as a biofilm matrix that protects the bacteria against host immune defenses and antibiotic treatment.
AB - Overproduction of the exopolysaccharide alginate contributes to the pathogenicity and antibiotic tolerance of Pseudomonas aeruginosa in chronic infections. The second messenger, c-di-GMP, is a positive regulator of the production of various biofilm matrix components and is known to regulate alginate synthesis at the posttranslational level in P. aeruginosa. We provide evidence that c-di-GMP also regulates transcription of the alginate operon in P. aeruginosa. Previous work has shown that transcription of the alginate operon is regulated by nine different proteins, AmrZ, AlgP, IHF alpha, IHF beta, CysB, Vfr, AlgR, AlgB, and AlgQ, and we investigated if some of these proteins function as a c-di-GMP effector. We found that deletion of algP, algQ, IHF alpha, and IHF beta had only a marginal effect on the transcription of the alginate operon. Deletion of vfr and cysB led to decreased transcription of the alginate operon, and the dependence of the c-di-GMP level was less pronounced, indicating that Vfr and CysB could be partially required for c-di-GMP-mediated regulation of alginate operon transcription. Our experiments indicated that the AmrZ, AlgR, and AlgB proteins are absolutely required for transcription of the alginate operon. However, differential radial capillary action of ligand assay (DRaCALA) and site-directed mutagenesis indicated that c-di-GMP does not bind to any of the AmrZ, AlgR, and AlgB proteins. IMPORTANCE The proliferation of alginate-overproducing P. aeruginosa variants in the lungs of cystic fibrosis patients often leads to chronic infection. The alginate functions as a biofilm matrix that protects the bacteria against host immune defenses and antibiotic treatment. Knowledge about the regulation of alginate synthesis is important in order to identify drug targets for the development of medicine against chronic P. aeruginosa infections. We provide evidence that c-di-GMP positively regulates transcription of the alginate operon in P. aeruginosa. Moreover, we revisited the role of the known alginate regulators, AmrZ, AlgP, IHF alpha, IHF beta, CysB, Vfr, AlgR, AlgB, and AlgQ, and found that their effect on transcription of the alginate operon is highly varied. Deletion of algP, algQ, IHF alpha, or IHF beta only had a marginal effect on transcription of the alginate operon, whereas deletion of vfr or cysB led to decreased transcription and deletion of amrZ, algR, or algB abrogated transcription.The proliferation of alginate-overproducing P. aeruginosa variants in the lungs of cystic fibrosis patients often leads to chronic infection. The alginate functions as a biofilm matrix that protects the bacteria against host immune defenses and antibiotic treatment.
KW - c-di-GMP
KW - alginate
KW - P
KW - aeruginosa
KW - transcriptional regulators
KW - BINDING PROTEIN
KW - BIOFILM FORMATION
KW - ALGD PROMOTER
KW - GENE
KW - EXOPOLYSACCHARIDE
KW - DNA
KW - RECEPTOR
KW - DOMAIN
KW - IDENTIFICATION
KW - SYSTEM
U2 - 10.1128/spectrum.00675-22
DO - 10.1128/spectrum.00675-22
M3 - Journal article
C2 - 35862969
VL - 10
JO - Microbiology spectrum
JF - Microbiology spectrum
SN - 2165-0497
IS - 4
M1 - e00675-22
ER -
ID: 315981728