Septal Class A Penicillin-Binding Protein Activity and ld-Transpeptidases Mediate Selection of Colistin-Resistant Lipooligosaccharide-Deficient Acinetobacter baumannii

• Published in: mBio 2021-01, Vol.12 (1)
• Main Authors: Kang, Katie N, Kazi, Misha I, Biboy, Jacob, Gray, Joe, Bovermann, Hannah, Ausman, Jessie, Boutte, Cara C, Vollmer, Waldemar, Boll, Joseph M
• Format: Article
• Language: English
• Subjects: Acinetobacter baumannii - genetics; Acinetobacter baumannii - isolation & purification; Acinetobacter baumannii - metabolism; Acinetobacter Infections - microbiology; Anti-Bacterial Agents - pharmacology; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Cell Membrane - metabolism; Colistin - pharmacology; Drug Resistance, Multiple, Bacterial - genetics; Gene Expression Regulation, Bacterial - drug effects; Humans; Lipid A - metabolism; Lipopolysaccharides - deficiency; Lipopolysaccharides - genetics; Microbial Sensitivity Tests; Molecular Biology and Physiology; Penicillin-Binding Proteins - metabolism; Peptidoglycan Glycosyltransferase; Peptidyl Transferases - metabolism; Research Article
• ISSN: 2161-2129; 2150-7511
• DOI: 10.1128/mBio.02185-20

Despite dogma suggesting that lipopolysaccharide/lipooligosaccharide (LOS) was essential for viability of Gram-negative bacteria, several clinical isolates produced LOS colonies after colistin selection. Inactivation of the conserved class A penicillin-binding protein, PBP1A, was a compensatory mutation that supported isolation of LOS , but the impact of PBP1A mutation was not characterized. Here, we show that the absence of PBP1A causes septation defects and that these, together with ld-transpeptidase activity, support isolation of LOS PBP1A contributes to proper cell division in , and its absence induced cell chaining. Only isolates producing three or more septa supported selection of colistin-resistant LOS PBP1A was enriched at the midcell, where the divisome complex facilitates daughter cell formation, and its localization was dependent on glycosyltransferase activity. Transposon mutagenesis showed that genes encoding two putative ld-transpeptidases (LdtJ and LdtK) became essential in the PBP1A mutant. Both LdtJ and LdtK were required for selection of LOS , but each had distinct enzymatic activities in the cell. Together, these findings demonstrate that defects in PBP1A glycosyltransferase activity and ld-transpeptidase activity remodel the cell envelope to support selection of colistin-resistant LOS The increasing prevalence of antibiotic treatment failure associated with Gram-negative bacterial infections highlights an urgent need to develop new alternative therapeutic strategies. The last-line antimicrobial colistin (polymyxin E) targets the ubiquitous outer membrane lipopolysaccharide (LPS)/LOS membrane anchor, lipid A, which is essential for viability of most diderms. However, several LOS clinical isolates were recovered after colistin selection, suggesting a conserved resistance mechanism. Here, we characterized a role for penicillin-binding protein 1A in septation and intrinsic β-lactam susceptibility. We also showed that defects in PBP1A glycosyltransferase activity and ld-transpeptidase activity support isolation of colistin-resistant LOS .