FimL regulates cAMP synthesis in Pseudomonas aeruginosa.
Pseudomonas aeruginosa, a ubiquitous bacteria found in diverse ecological niches, is an important cause of acute infections in immunocompromised individuals and chronic infections in patients with Cystic Fibrosis. One signaling molecule required for the coordinate regulation of virulence factors associated with acute infections is 3', 5'-cyclic adenosine monophosphate, (cAMP), which binds to and activates a catabolite repressor homolog, Vfr. Vfr controls the transcription of many virulence factors, including those associated with Type IV pili (TFP), the Type III secretion system (T3SS), the Type II secretion system, flagellar-mediated motility, and quorum sensing systems. We previously identified FimL, a protein with histidine phosphotransfer-like domains, as a regulator of Vfr-dependent processes, including TFP-dependent motility and T3SS function. In this study, we carried out genetic and physiologic studies to further define the mechanism of action of FimL. Through a genetic screen designed to identify suppressors of FimL, we found a putative cAMP-specific phosphodiesterase (CpdA), suggesting that FimL regulates cAMP levels. Inactivation of CpdA increases cAMP levels and restores TFP-dependent motility and T3SS function to fimL mutants, consistent with in vivo phosphodiesterase activity. By constructing combinations of double and triple mutants in the two adenylate cyclase genes (cyaA and cyaB), fimL, and cpdA, we show that ΔfimL mutants resemble ΔcyaB mutants in TM defects, decreased T3SS transcription, and decreased cAMP levels. Similar to some of the virulence factors that they regulate, we demonstrate that CyaB and FimL are polarly localized. These results reveal new complexities in the regulation of diverse virulence pathways associated with acute P. aeruginosa infections.
Department of Medicine, University of California San Francisco, San Francisco, California, United States of America.
Yuki F
YF
Inclan
Medora J
MJ
Huseby
Joanne N
JN
Engel
D037361:Adenylate Cyclase Toxin; D000262:Adenylyl Cyclases / Q000235:genetics; D018829:Adhesins, Bacterial / Q000235:genetics / Q000378:metabolism*; D000242:Cyclic AMP / Q000096:biosynthesis*; D036781:Fimbriae Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009154:Mutation; D011552:Pseudomonas Infections; D011550:Pseudomonas aeruginosa / Q000378:metabolism* / Q000472:pathogenicity; D037521:Virulence Factors
D037361:Adenylate Cyclase Toxin; D018829:Adhesins, Bacterial; D037521:Virulence Factors; D036781:Fimbriae Proteins; D000242:Cyclic AMP; D000262:Adenylyl Cyclases
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