Biology subjects

Palmer, L. D.

Publications and source records attributed to Palmer, L. D..

4 recordsLinked to original sources

bioRxiv2025

A regulator of amino acid catabolism controls Acinetobacter baumannii gut colonization

Asymptomatic gut colonization increases the risk of clinical infection and transmission by the multidrug-resistant pathogen Acinetobacter baumannii. Ornithine utilization was shown to be critical for A. baumannii competition with the resident microbiota to persist in gut colonization, but the regulatory mechanisms and cues are unknown. Here, we identify a transcriptional regulator, AstR, that specifically activates the expression of the A. baumannii ornithine utilization operon astNOP. Phylogenetic analysis suggests that AstR was co-opted from the Acinetobacter arginine utilization ast(G)CADBE locus and is specialized to regulate ornithine utilization in A. baumannii. Reporter assays showed that astN promoter expression was activated by ornithine but inhibited by glutamate and other preferred amino acids. astN promoter expression was similarly activated by incubation with fecal samples from conventional mice but not germ-free mice, suggesting AstR-dependent activation of the astN promoter responds to intermicrobial competition for amino acids. Finally, AstR was required for A. baumannii to colonize the gut in a mouse model. Together, these results suggest that pathogenic Acinetobacter species evolved AstR to regulate ornithine catabolism, which is required to compete with the microbiota during gut colonization.

microbiology↗

bioRxiv2025

DcaP-Family Porins are Required for Carboxylic Acid Catabolism in Acinetobacter baumannii

Acinetobacter baumannii is a pathogen of concern and a leading cause of multidrug-resistant healthcare-associated infections. The A. baumannii outer membrane is a barrier to antimicrobials and host defenses but must also allow essential nutrients to permeate. Here, we investigate the functional importance of the putative DcaP-family outer membrane porins, which include a proposed vaccine target in A. baumannii. All A. baumannii genomes surveyed encode multiple DcaP family porins, which we classify in four classes based on protein sequence phylogeny (DcaP1-4). DcaP proteins encoded by species in other genera could not be mapped to these DcaP classes and phylogenetic analysis suggests DcaP1-4 proteins diversified within Acinetobacter. Phenotypic array assays and additional experiments show that the DcaP proteins were necessary for growth on multiple di- and tri-carboxylic acids as sole carbon sources, including citric acid and tricarballylic acid. Specifically, DcaP3 was required; however, DcaP1, DcaP2, and DcaP4 were all able to complement the total dcaP knockout to varying degrees, suggesting partial functional overlap. Finally, a mutant lacking all DcaP proteins was attenuated in the liver and spleen in a mouse model of bloodstream infection and complemented by expression of DcaP3. However, a {Delta}dcaP3 mutant had no defect, demonstrating functional redundancy among DcaP proteins during infection. These findings provide insight on how A. baumannii acquires nutrients through the outer membrane barrier and show DcaP proteins are important during infection in specific host niches, validating their potential as a therapeutic target.

microbiology↗

bioRxiv2024

Amino acid competition shapes Acinetobacter baumannii gut carriage

Antimicrobial resistance is an urgent threat to human health. Asymptomatic colonization is often critical for persistence of antimicrobial-resistant pathogens. Gut colonization by the antimicrobial-resistant priority pathogen Acinetobacter baumannii is associated with increased risk of clinical infection. Ecological factors shaping A. baumannii gut colonization remain unclear. Here we show that A. baumannii and other pathogenic Acinetobacter evolved to utilize the amino acid ornithine, a non- preferred carbon source. A. baumannii utilizes ornithine to compete with the resident microbiota and persist in the gut in mice. Supplemental dietary ornithine promotes long-term fecal shedding of A. baumannii. By contrast, supplementation of a preferred carbon source--monosodium glutamate (MSG)-- abolishes the requirement for A. baumannii ornithine catabolism. Additionally, we report evidence for diet promoting A. baumannii gut carriage in humans. Together, these results highlight that evolution of ornithine catabolism allows A. baumannii to compete with the microbiota in the gut, a reservoir for pathogen spread.

microbiology↗

bioRxiv2023

Genetic synergy in Acinetobacter baumannii undecaprenyl biosynthesis and maintenance of lipid asymmetry impacts outer membrane and antibiotic resistance

Acinetobacter baumannii is a Gram-negative healthcare-associated pathogen that poses a major health concern due to increasing multidrug resistance. The Gram-negative cell envelope is a key barrier to antimicrobial entry and includes an inner and outer membrane. The outer membrane has an asymmetric composition that is important for structural integrity and barrier to the environment. Therefore, Gram-negative bacteria have mechanisms to uphold this asymmetry such as the maintenance of lipid asymmetry system (Mla), which removes glycerophospholipids from the outer leaflet of the outer membrane and transports them to the inner membrane. Loss of this system in A. baumannii results in attenuated virulence and increased susceptibility to membrane stressors and some antibiotics. We recently reported two strain variants of the A. baumannii type strain ATCC 17978, 17978VU and 17978UN. We show here that {Delta}mlaF mutants in the two strains display different phenotypes for membrane stress resistance, antibiotic resistance, and pathogenicity in a murine pneumonia model. We used comparative genetics to identify interactions between ATCC 17978 strain alleles and mlaF to uncover the cause behind the phenotypic differences. Although allele differences in obgE were previously reported to synergize with {Delta}mlaF to affect growth and stringent response, we show that obgE alleles do not affect membrane stress resistance. Instead, a single nucleotide polymorphism (SNP) in the essential gene encoding undecaprenyl pyrophosphate (Und-PP) synthase, uppS, synergizes with {Delta}mlaF to increase susceptibility to membrane stress and antibiotics, and reduce persistence in a mouse lung infection. Und-P is a lipid glycan carrier known to be required for biosynthesis of A. baumannii capsule, cell wall, and glycoproteins. Our data suggest that in the absence of the Mla system, the cellular level of Und-P is critical for envelope integrity, antibiotic resistance, and lipooligosaccharide abundance. These findings uncover synergy between Und-P and the Mla system in maintaining the A. baumannii outer membrane and stress resistance.

microbiology↗