title: Nitazoxanide Inhibits Pilus Biogenesis by Interfering with Folding of the Usher Protein in the Outer Membrane.
abstract: Many bacterial pathogens assemble surface fibers termed pili or fimbriae that facilitate attachment to host cells and colonization of host tissues. The chaperone/usher (CU) pathway is a conserved secretion system that is responsible for the assembly of virulence-associated pili by many different Gram-negative bacteria. Pilus biogenesis by the CU pathway requires a dedicated periplasmic chaperone and an integral outer membrane (OM) assembly and secretion platform termed the usher. Nitazoxanide (NTZ), an antiparasitic drug, was previously shown to inhibit the function of aggregative adherence fimbriae and type 1 pili assembled by the CU pathway in enteroaggregativeEscherichia coli, an important causative agent of diarrhea. We show here that NTZ also inhibits the function of type 1 and P pili from uropathogenicE. coli(UPEC). UPEC is the primary causative agent of urinary tract infections, and type 1 and P pili mediate colonization of the bladder and kidneys, respectively. By analysis of the different stages of the CU pilus biogenesis pathway, we show that treatment of bacteria with NTZ causes a reduction in the number of usher molecules in the OM, resulting in a loss of pilus assembly on the bacterial surface. In addition, we determine that NTZ specifically prevents proper folding of the usher β-barrel domain in the OM. Our findings demonstrate that NTZ is a pilicide with a novel mechanism of action and activity against diverse CU pathways. This suggests that further development of the NTZ scaffold may lead to new antivirulence agents that target the usher to prevent pilus assembly.
authors.affiliation: Center for Infectious Diseases and Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Chahales
authors.affiliation: Department of Medicine, Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, Virginia, USA.
authors.forename: Paul S
authors.initials: PS
authors.lastname: Hoffman
authors.affiliation: Center for Infectious Diseases and Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA david.thanassi@stonybrook.edu.
authors.forename: David G
authors.initials: DG
authors.lastname: Thanassi
mesh_terms: D000818:Animals; D000977:Antiparasitic Agents / Q000494:pharmacology*; D058947:Bacterial Secretion Systems / Q000187:drug effects; D003001:Cloning, Molecular; D004912:Erythrocytes / Q000187:drug effects; D029968:Escherichia coli Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010861:Fimbriae, Bacterial / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006168:Guinea Pigs; D006386:Hemagglutination Tests; D006801:Humans; D018832:Molecular Chaperones / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009574:Nitro Compounds; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D000072757:Protein Conformation, beta-Strand; D017510:Protein Folding / Q000187:drug effects; D055503:Protein Multimerization / Q000187:drug effects; D021122:Protein Subunits / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013844:Thiazoles / Q000494:pharmacology*; D056689:Uropathogenic Escherichia coli / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics / Q000378:metabolism
chemical_list: D000977:Antiparasitic Agents; D058947:Bacterial Secretion Systems; C574723:CfaC protein, E coli; D029968:Escherichia coli Proteins; D018832:Molecular Chaperones; D009574:Nitro Compounds; D021122:Protein Subunits; D013844:Thiazoles; C041747:nitazoxanide
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