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PXD029140-1

PXD029140 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleImpacts of resistance gene knock-outs on the proteome
DescriptionThe spread of antibiotic resistance has developed to all known antibiotics. Extended spectrum β-lactamase-producing bacteria are particularly problematic, as they are resistant to a wide range of commonly used antibiotics. Resistance to β-lactams is known to be multifactorial, although the underlying mechanisms generally are poorly understood but critical factors for effective therapy against infections, especially for multi-resistant pathogenic bacteria. In the present study, a plasmid-based homologous recombination system was used to target and delete specific β-lactamase genes (i.e., the blaOXA-1, blaTEM-1 or the ESBL blaCTX-M15) of the clinical strain ESBL Escherichia coli CCUG 73778, generating three “knock-out” clone variants, each one lacking only one of the β-lactamases. The objective was to determine the genotypic impacts of each gene loss on the phenotypic antibiotic resistance and proteome of the bacterium. Quantitative proteomic analyses performed on the three clone variants and the original strain, using tandem mass tags (TMT) and bottom-up liquid chromatography tandem mass spectrometry (LC-MS/MS), after exposure to different concentrations of cefadroxil. Variation of the proteome in each clone variant was determined, to establish the relative importance of each resistance gene and better understand the genetic and proteomic responses and mechanisms of the resistance phenotypes. The knockout of blaCTX-M-15 was observed to have the greatest impact in protein expression, with the knockout of blaOXA-1 also effecting a marked but lower degree of changes. Proteins known to be associated with antibiotic resistance, cell membrane integrity, cellular stress, gene expression and hypothetical/unknown function proteins, among others, demonstrated distinct differences in expression levels (Fold change >1-5 or <-1.5), that may be related to aspects of compensation for the mutant resistance phenotypes. The present study provides a framework to study the impacts of targeted loss of antibiotic resistance genes in clinically relevant strains for understanding the mechanisms of phenotypic antibiotic resistance.
HostingRepositoryPRIDE
AnnounceDate2023-01-04
AnnouncementXMLSubmission_2023-01-04_04:53:49.434.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterProteomicsCore Facility
SpeciesList scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationListTMT6plex-126 reporter+balance reagent acylated residue; monohydroxylated residue
InstrumentOrbitrap Fusion Lumos
Dataset History
RevisionDatetimeStatusChangeLog Entry
02021-10-15 03:21:40ID requested
12023-01-04 04:53:49announced
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: Escherichia coli,antibiotic resistance, β-lactamase, quantitative proteomics, tandem mass tag, LC-MS/MS
Contact List
EdwardMoore
contact affiliationProfessor,Department of Infectious Diseases, University of Gothenburg; Box 480, 40530 Göteborg, Sweden
contact emailedward.moore@gu.se
lab head
ProteomicsCore Facility
contact affiliationSAMBIO Core Facilities, Sahgrenska Academy, University of Gothenburg
contact emailgupcf@outlook.com
dataset submitter
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Dataset FTP location
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