Characterization of UDP-glucose dehydrogenase and UDP-glucose pyrophosphorylase mutants of Proteus mirabilis: defectiveness in polymyxin B resistance, swarming, and virulence.
Proteus mirabilis is known to be highly resistant to the action of polymyxin B (PB). However, the mechanism underlying PB resistance is not clear. In this study, we used Tn5 transposon mutagenesis to identify genes that may affect PB resistance in P. mirabilis. Two genes, ugd and galU, which may encode UDP-glucose dehydrogenase (Ugd) and UDP-glucose pyrophosphorylase (GalU), respectively, were identified. Knockout mutants of ugd and galU were found to be extremely sensitive to PB, presumably because of alterations in lipopolysaccharide (LPS) structure and cell surface architecture in these mutants. These mutants were defective in swarming, expressed lower levels of virulence factor hemolysin, and had lower cell invasion ability. Complementation of the ugd or galU mutant with the full-length ugd or galU gene, respectively, led to the restoration of wild-type phenotypic traits. Interestingly, we found that the expression of Ugd and GalU was induced by PB through RppA, a putative response regulator of the bacterial two-component system that we identified previously. Mutation in either ugd or galU led to activation of RpoE, an extracytoplasmic function sigma factor that has been shown to be activated by protein misfolding and alterations in cell surface structure in other bacteria. Activation of RpoE or RpoE overexpression was found to cause inhibition of FlhDC and hemolysin expression. To our knowledge, this is the first report describing the roles and regulation of Ugd and GalU in P. mirabilis.
Jiang
Sin-Sien
SS
Department and Graduate Institute of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine, National Taiwan University, Taipei, Taiwan, ROC.
Lin
Tzu-Yi
TY
Wang
Won-Bo
WB
Liu
Ming-Che
MC
Hsueh
Po-Ren
PR
Liaw
Shwu-Jen
SJ
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000378:metabolism; D002473:Cell Wall / Q000187:drug effects; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D005408:Flagellin / Q000235:genetics; D024841:Fluoroquinolones / Q000494:pharmacology*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D055786:Gene Knockout Techniques; D006460:Hemolysin Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009068:Movement / Q000502:physiology; D016296:Mutagenesis; D010641:Phenotype; D011112:Polymyxin B / Q000494:pharmacology*; D011513:Proteus mirabilis / Q000187:drug effects / Q000235:genetics* / Q000472:pathogenicity; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000235:genetics* / Q000378:metabolism; D014533:Uridine Diphosphate Glucose Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D014774:Virulence
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D024841:Fluoroquinolones; D006460:Hemolysin Proteins; C412036:RppA protein, Synechocystis; D005408:Flagellin; D014533:Uridine Diphosphate Glucose Dehydrogenase; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase; D011112:Polymyxin B