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Zinc limitation in Klebsiella pnuemoniae

ABSTRACT: Nutrient adaptation is key in limiting environments for the promotion of microbial growth and survival. In microbial systems, zinc is an important component for many cellular processes and bioavailability varies greatly among different conditions. In the bacterium, Klebsiella pneumoniae, the impact of zinc limitation has not been explored at the protein level. Here, we apply a mass spectrometry-based quantitative proteomics strategy to profile the global impact of zinc limitation on the cellular proteome and extracellular environment (secretome) of K. pneumoniae. Our data defines the impact of zinc on proteins involved in transcriptional regulation and emphasizes the modulation of a vast array of proteins associated with zinc acquisition, transport, and binding. We also identify proteins in the extracellular environment associated with conventional and nonconventional modes of secretion. In particular, we focus on the impact of zinc on an uncharacterized cation transporter and the role of the ChaB transporter in influencing bacterial growth, capsule production, nad virulence. Moreover, we validate our results through transcriptional profiling of genes reprsessed under high abudnance of a histidine repression operon, hutC. Overall, we provide evidence of novel connections between zinc availability and capsule production in a bacterial system and define new roles for a transporter.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Klebsiella Pneumoniae

SUBMITTER: Jennifer Geddes-McAlister

LAB HEAD: Jennifer Geddes-McAlister

PROVIDER: PXD016042 | Pride | 2021-02-12

REPOSITORIES: Pride

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Project description:The emergence of hypervirulent Klebsiella pneumoniae (hvKP) strains poses a significant threat to public health due to their high mortality rates and propensity to cause severe community-acquired infections in otherwise healthy individuals. The ability of hvKP to form biofilms and produce a protective capsule contributes to its enhanced virulence and is a significant challenge to effective antibiotic treatment. Therefore, understanding the molecular mechanisms underlying hvKP virulence and biofilm formation is crucial for developing new therapeutic strategies. Polyphosphate Kinase 1 (PPK1) is an enzyme responsible for inorganic polyphosphate synthesis and plays a vital role in regulating various physiological processes in bacteria. In this study, we investigated the impact of polyP metabolism on the biofilm and capsule formation and virulence traits in hvKP using Dictyostelium discoideum amoeba as a model host. We found that the PPK1 null-mutant was impaired in biofilm and capsule formation and showed attenuated virulence in D. discoideum compared to the wild-type strain. We performed a shotgun proteomic analysis of the PPK1 mutant and wild-type strain to gain further insight into the underlying molecular mechanism. The results revealed that the PPK1 mutant had a differential expression of proteins (DEP) involved in capsule synthesis (Wzi - Ugd), biofilm formation (MrkC-D-H), synthesis of the colibactin genotoxin precursor (ClbB), as well as proteins associated with the synthesis and modification of lipid A (ArnB -LpxC - PagP). These proteomic findings corroborate the phenotypic observations and indicate that the PPK1 mutation is associated with impaired biofilm and capsule formation and attenuated virulence in hypervirulent K. pneumoniae. Overall, our study highlights the importance of polyP synthesis in regulating extracellular biomolecules and virulence in K. pneumoniae and provides insights into potential therapeutic targets for treating K. pneumoniae infections.

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Zinc limitation in Klebsiella pnuemoniae

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