Journal Article

An inhibitor/anti-inhibitor system controls the activity of lytic transglycosylase MltF in Pseudomonas aeruginosa

Wang, Michelle, Darwin, Andrew J., Xiao Ma, Sheya

2023

Overview

Most bacterial cell envelopes contain a cell wall layer made of peptidoglycan. The synthesis of new peptidoglycan is critical for cell growth, division, and morphogenesis and is also coordinated with peptidoglycan hydrolysis to accommodate the new material. However, the enzymes that cleave peptidoglycan must be carefully controlled to avoid autolysis. In recent years, some control mechanisms have begun to emerge, although there are many more questions than answers for how most cell wall hydrolases are regulated. Here, we report a novel cell wall hydrolase control mechanism in Pseudomonas aeruginosa , which we discovered during our characterization of a mutant sensitive to the overproduction of a secretin protein. The mutation affected an uncharacterized Sel1-like repeat protein encoded by the PA3978 locus. In addition to the secretin-sensitivity phenotype, PA3978 disruption also increased resistance to a β-lactam antibiotic used in the clinic. In vivo and in vitro analyses revealed that PA3978 binds to the catalytic domain of the lytic transglycosylase MltF and inhibits its activity. ∆PA3978 mutant phenotypes were suppressed by deleting mltF , consistent with them having been caused by elevated MltF activity. We also discovered another interaction partner of PA3978 encoded by the PA5502 locus. The phenotypes of a ∆PA5502 mutant suggested that PA5502 interferes with the inhibitory function of PA3978 toward MltF, and we confirmed that activity for PA5502 in vitro . Therefore, PA3978 and PA5502 form an inhibitor/anti-inhibitor system that controls MltF activity. We propose to name these proteins IltA ( i nhibitor A of l ytic t ransglycosylase) and LiiA ( l ytic transglycosylase i nhibitor A’s i nhibitor). A peptidoglycan cell wall is an essential component of almost all bacterial cell envelopes, which determines cell shape and prevents osmotic rupture. Antibiotics that interfere with peptidoglycan synthesis have been one of the most important treatments for bacterial infections. Peptidoglycan must also be hydrolyzed to incorporate new material for cell growth and division and to help accommodate important envelope-spanning systems. However, the enzymes that hydrolyze peptidoglycan must be carefully controlled to prevent autolysis. Exactly how this control is achieved is poorly understood in most cases but is a highly active area of current research. Identifying hydrolase control mechanisms has the potential to provide new targets for therapeutic intervention. The work here reports the important discovery of a novel inhibitor/anti-inhibitor system that controls the activity of a cell wall hydrolase in the human pathogen Pseudomonas aeruginosa , which also affects resistance to an antibiotic used in the clinic.

• Publisher: American Society for Microbiology
• Subject: Anti-Bacterial Agents - pharmacology / Antibiotics / Autolysis / Bacterial Proteins - genetics / Bacterial Proteins - metabolism / Bacteriology / cell envelope / Cell envelopes / Cell growth / Cell size / cell wall / Cell Wall - drug effects / Cell Wall - metabolism / Cell walls / Enzyme Inhibitors - pharmacology / enzyme regulation / Experiments / Genes / Genotype & phenotype / Glycosyltransferases - antagonists & inhibitors / Glycosyltransferases - genetics / Glycosyltransferases - metabolism / Humans / Hydrolase / Mass spectrometry / Metabolism / Morphogenesis / Multidrug resistant organisms / Mutants / Mutation / Nosocomial infections / Peptides / Peptidoglycan - metabolism / Peptidoglycans / Phenotypes / Proteins / Pseudomonas aeruginosa / Pseudomonas aeruginosa - drug effects / Pseudomonas aeruginosa - enzymology / Pseudomonas aeruginosa - genetics / Research Article / Scientific imaging / Secretin / Toxicity / Virulence / β-Lactam antibiotics