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Polyphosphate kinase regulates LPS structure and polymyxin resistance during starvation in E. coli.
Baijal, Kanchi; Abramchuk, Iryna; Herrera, Carmen M; Mah, Thien-Fah; Trent, M Stephen; Lavallée-Adam, Mathieu; Downey, Michael.
Afiliação
  • Baijal K; Ottawa Institute of Systems Biology, Ottawa, Ontario, Canada.
  • Abramchuk I; Department of Cellular & Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada.
  • Herrera CM; Ottawa Institute of Systems Biology, Ottawa, Ontario, Canada.
  • Mah TF; Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, Ontario, Canada.
  • Trent MS; Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, Georgia, United States of America.
  • Lavallée-Adam M; Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, Ontario, Canada.
  • Downey M; Centre for Infection, Immunity, and Inflammation, University of Ottawa, Ottawa, Ontario, Canada.
PLoS Biol ; 22(3): e3002558, 2024 Mar.
Article em En | MEDLINE | ID: mdl-38478588
ABSTRACT
Polyphosphates (polyP) are chains of inorganic phosphates that can reach over 1,000 residues in length. In Escherichia coli, polyP is produced by the polyP kinase (PPK) and is thought to play a protective role during the response to cellular stress. However, the molecular pathways impacted by PPK activity and polyP accumulation remain poorly characterized. In this work, we used label-free mass spectrometry to study the response of bacteria that cannot produce polyP (Δppk) during starvation to identify novel pathways regulated by PPK. In response to starvation, we found 92 proteins significantly differentially expressed between wild-type and Δppk mutant cells. Wild-type cells were enriched for proteins related to amino acid biosynthesis and transport, while Δppk mutants were enriched for proteins related to translation and ribosome biogenesis, suggesting that without PPK, cells remain inappropriately primed for growth even in the absence of the required building blocks. From our data set, we were particularly interested in Arn and EptA proteins, which were down-regulated in Δppk mutants compared to wild-type controls, because they play a role in lipid A modifications linked to polymyxin resistance. Using western blotting, we confirm differential expression of these and related proteins in K-12 strains and a uropathogenic isolate, and provide evidence that this mis-regulation in Δppk cells stems from a failure to induce the BasRS two-component system during starvation. We also show that Δppk mutants unable to up-regulate Arn and EptA expression lack the respective L-Ara4N and pEtN modifications on lipid A. In line with this observation, loss of ppk restores polymyxin sensitivity in resistant strains carrying a constitutively active basR allele. Overall, we show a new role for PPK in lipid A modification during starvation and provide a rationale for targeting PPK to sensitize bacteria towards polymyxin treatment. We further anticipate that our proteomics work will provide an important resource for researchers interested in the diverse pathways impacted by PPK.
Assuntos

Texto completo: 1 Bases de dados: MEDLINE Assunto principal: Lipopolissacarídeos / Fosfotransferases (Aceptor do Grupo Fosfato) / Escherichia coli Idioma: En Revista: PLoS Biol Assunto da revista: BIOLOGIA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Canadá

Texto completo: 1 Bases de dados: MEDLINE Assunto principal: Lipopolissacarídeos / Fosfotransferases (Aceptor do Grupo Fosfato) / Escherichia coli Idioma: En Revista: PLoS Biol Assunto da revista: BIOLOGIA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Canadá