Cyclic-di-GMP-mediated repression of swarming motility by Pseudomonas aeruginosa: the pilY1 gene and its impact on surface-associated behaviors.
The intracellular signaling molecule cyclic-di-GMP (c-di-GMP) has been shown to influence surface-associated behaviors of Pseudomonas aeruginosa, including biofilm formation and swarming motility. Previously, we reported a role for the bifA gene in the inverse regulation of biofilm formation and swarming motility. The bifA gene encodes a c-di-GMP-degrading phosphodiesterase (PDE), and the Delta bifA mutant exhibits increased cellular pools of c-di-GMP, forms hyperbiofilms, and is unable to swarm. In this study, we isolated suppressors of the Delta bifA swarming defect. Strains with mutations in the pilY1 gene, but not in the pilin subunit pilA gene, show robust suppression of the swarming defect of the Delta bifA mutant, as well as its hyperbiofilm phenotype. Despite the ability of the pilY1 mutation to suppress all the c-di-GMP-related phenotypes, the global pools of c-di-GMP are not detectably altered in the Delta bifA Delta pilY1 mutant relative to the Delta bifA single mutant. We also show that enhanced expression of the pilY1 gene inhibits swarming motility, and we identify residues in the putative VWA domain of PilY1 that are important for this phenotype. Furthermore, swarming repression by PilY1 specifically requires the diguanylate cyclase (DGC) SadC, and epistasis analysis indicates that PilY1 functions upstream of SadC. Our data indicate that PilY1 participates in multiple surface behaviors of P. aeruginosa, and we propose that PilY1 may act via regulation of SadC DGC activity but independently of altering global c-di-GMP levels.
Department of Microbiology and Immunology, Dartmouth Medical School, North College Street, Hanover, NH 03755, USA.
S L
SL
Kuchma
A E
AE
Ballok
J H
JH
Merritt
J H
JH
Hammond
W
W
Lu
J D
JD
Rabinowitz
George A
GA
O'Toole
D000595:Amino Acid Sequence; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000378:metabolism; D036781:Fimbriae Proteins / Q000235:genetics / Q000378:metabolism*; D010861:Fimbriae, Bacterial / Q000502:physiology; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D005838:Genotype; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000378:metabolism*
C103855:PilY1 protein, Pseudomonas aeruginosa; D036781:Fimbriae Proteins; C062025:bis(3',5')-cyclic diguanylic acid; D006152:Cyclic GMP