PXD043566 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Proteomics analysis reveals a role for E. coli polyphosphate kinase in regulation of membrane structure and polymyxin resistance via the BasS-BasR two component system |
Description | Polyphosphates (polyP) are chains of inorganic phosphates that can reach 1000s of 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 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 (∆ppk mutants) during nutrient starvation and to identify novel pathways regulated by PPK. In response to nutrient starvation, we found 92 proteins significantly differentially expressed between wild-type and ∆ppkmutant cells. Wild-type cells were enriched for proteins related to amino acid biosynthesis and transport while Δppkmutants were enriched for proteins related to translation and ribosome biogenesis, suggesting that without PPK cells remain inappropriately primed for growth. From our data set, we were particularly interested in Arn and EptA proteins, which were downregulated in ∆ppk 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, and provide evidence that this mis-regulation in ∆ppk cells stems from a failure to induce the BasS/BasR two component system during starvation. We also show that ∆ppk mutants unable to upregulate 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 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 diverse pathways impacted by PPK. |
HostingRepository | PRIDE |
AnnounceDate | 2024-02-14 |
AnnouncementXML | Submission_2024-02-14_11:29:01.871.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Iryna Abramchuk |
SpeciesList | scientific name: Escherichia coli; NCBI TaxID: 562; |
ModificationList | No PTMs are included in the dataset |
Instrument | Orbitrap Fusion Lumos |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2023-07-05 13:05:13 | ID requested | |
⏵ 1 | 2024-02-14 11:29:02 | announced | |
Publication List
Dataset with its publication pending |
Keyword List
submitter keyword: LC-MSMS, cell culture, bacteria, label-free quantification,E. coli |
Contact List
Michael Downey |
contact affiliation | Ottawa Institute of Systems Biology, Ottawa, Ontario, Canada Department of Cellular & Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada |
contact email | mdowne2@uottawa.ca |
lab head | |
Iryna Abramchuk |
contact affiliation | University of Ottawa |
contact email | iabra008@uottawa.ca |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD043566
- Label: PRIDE project
- Name: Proteomics analysis reveals a role for E. coli polyphosphate kinase in regulation of membrane structure and polymyxin resistance via the BasS-BasR two component system