PXD074122-1

PXD074122 is an original dataset announced via ProteomeXchange.

Title	The MATE transporter CD20030 modulates multidrug resistance and oxidative stress defense while orchestrating a motility-virulence trade-off via metabolic reprogramming in Clostridioides difficile
Description	Background: The multidrug-resistant pathogen Clostridioides difficile poses a severe public health threat. While Multidrug and Toxic Compound Extrusion (MATE) transporters are known to mediate xenobiotic efflux and oxidative stress responses in other bacterial species, their physiological roles in C. difficile—beyond the characterized CdeA pump—remain largely unexplored. Methods: This study functionally characterized a putative MATE transporter gene (mate, CD20030) in C. difficile strain 630. Utilizing a CRISPR-Cas9 genome editing system, we constructed a markerless deletion mutant (Δmate) and a complemented strain to systematically evaluate antimicrobial susceptibility, growth kinetics, autolysis, and oxidative stress tolerance. Furthermore, comparative proteomic profiling coupled with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were employed to unravel the molecular networks underlying the observed phenotypes. Results: The Δmate mutant exhibited broad-spectrum hypersensitivity to antibiotics (e.g., fluoroquinolones, β-lactams) and significantly reduced tolerance to hydrogen peroxide, confirming its dual role in drug efflux and oxidative stress defense. Intriguingly, mate deletion resulted in a pleiotropic phenotype characterized by delayed autolysis but a paradoxical upregulation of toxin genes (tcdA and tcdB), leading to enhanced cytotoxicity. Bioinformatic analyses (GO and KEGG) revealed that the loss of mate triggered extensive metabolic and structural reprogramming. Most notably, pathways associated with flagellar assembly, bacterial chemotaxis, and oxidoreductase activity were significantly downregulated, while metabolic pathways were upregulated. This suggests a strategic resource reallocation trade-off where energy is diverted from motility towards stress survival and virulence factor production. Conclusion: Our findings establish the mate gene as a central physiological hub in C. difficile, linking intrinsic multidrug resistance to oxidative stress adaptation, motility regulation, and virulence. This transporter functions not merely as a drug pump but as a critical determinant of cellular homeostasis and pathogenesis.
HostingRepository	iProX
AnnounceDate	2026-02-05
AnnouncementXML	Submission_2026-02-06_00:32:32.860.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Fahui Chen
SpeciesList	scientific name: Clostridioides difficile 630; NCBI TaxID: 272563;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Astral

Revision	Datetime	Status	ChangeLog Entry
0	2026-02-06 00:31:57	ID requested
⏵ 1	2026-02-06 00:32:33	announced

Dataset with its publication pending

submitter keyword: Clostridioides difficile, MATE efflux pump, CRISPR-Cas9, oxidative stress, virulence regulation, flagellar assembly, proteomics

Wei Hong
contact affiliation	Guizhou Medical University
contact email	hongwei_2015@hotmail.com
lab head
Fahui Chen
contact affiliation	Guizhou Medical University
contact email	1578134060@qq.com
dataset submitter

iProX dataset URI