Penicillin-Binding Protein 3 Is Essential for Growth of Pseudomonas aeruginosa.
Penicillin-binding proteins (PBPs) function as transpeptidases, carboxypeptidases, or endopeptidases during peptidoglycan synthesis in bacteria. As the well-known drug targets for β-lactam antibiotics, the physiological functions of PBPs and whether they are essential for growth are of significant interest. The pathogen Pseudomonas aeruginosa poses a particular risk to immunocompromised and cystic fibrosis patients, and infections caused by this pathogen are difficult to treat due to antibiotic resistance. To identify potential drug targets among the PBPs in P. aeruginosa, we performed gene knockouts of all the high-molecular-mass (HMM) PBPs and determined the impacts on cell growth and morphology, susceptibility to β-lactams, peptidoglycan structure, virulence, and pathogenicity. Disruptions of the transpeptidase domains of most HMM PBPs, including double disruptions, had only minimal effects on cell growth. The exception was PBP3, where cell growth occurred only when the protein was conditionally expressed on an integrated plasmid. Conditional deletion of PBP3 also caused a defect in cell division and increased susceptibility to β-lactams. Knockout of PBP1a led to impaired motility, and this observation, together with its localization at the cell poles, suggests its involvement in flagellar function. Overall, these findings reveal that PBP3 represents the most promising target for drug discovery against P. aeruginosa, whereas other HMM PBPs have less potential.
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina, USA.
Wei
W
Chen
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina, USA.
Yong-Mei
YM
Zhang
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina, USA davies@musc.edu.
Christopher
C
Davies
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D046915:Penicillin-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D010457:Peptidoglycan / Q000494:pharmacology; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D047090:beta-Lactams / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D046915:Penicillin-Binding Proteins; D010457:Peptidoglycan; D047090:beta-Lactams
Pseudomonas aeruginosa; antimicrobial drug targets; essential genes; genetic knockout; penicillin-binding proteins; peptidoglycan