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Quantitative SWATH Proteomic Analysis of Campylobacter jejuni and Campylobacter coli Following Exposure to the Antimicrobial Ionophore PBT2

ABSTRACT: Campylobacter jejuni ATCC 33560 and Campylobacter coli ATCC BAA-1061 were exposed to the antimicrobial ionophore PBT2 to investigate proteome-wide responses associated with its antibacterial activity. Cultures were grown in Mueller-Hinton broth supplemented with 5% defibrinated horse blood and harvested at defined time points following treatment with PBT2 or untreated control conditions. Quantitative proteomic profiling was performed using SWATH data-independent acquisition (DIA) mass spectrometry. The resulting dataset provides a comprehensive characterization of protein abundance changes associated with PBT2 exposure, including alterations in pathways related to translation, respiratory metabolism, cell envelope integrity, motility, and metal homeostasis. These data provide insights into the molecular mechanisms underlying the antimicrobial activity of PBT2 against clinically relevant Campylobacter species.

INSTRUMENT(S):

ORGANISM(S): Campylobacter Coli Rm2228 Campylobacter Jejuni Subsp. Jejuni Atcc 33560

DISEASE(S): Gastroenteritis

SUBMITTER: Huiting Chen

LAB HEAD: Freda Jen

PROVIDER: PXD080115 | Pride | 2026-06-27

REPOSITORIES: Pride

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Project description:Campylobacter jejuni is currently the leading cause of bacterial gastroenteritis in humans. Comparison of multiple Campylobacter strains revealed a high genetic and phenotypic diversity. However, little is known about differences in transcriptome organization, gene expression, and small RNA (sRNA) repertoires. Here we present the first comparative primary transcriptome analysis based on the differential RNAM-bM-^@M-^Sseq (dRNAM-bM-^@M-^Sseq) of four C. jejuni isolates. Our approach includes a novel, generic method for the automated annotation of transcriptional start sites (TSS), which allowed us to provide genome-wide promoter maps in the analyzed strains. These global TSS maps are refined through the integration of a SuperGenome approach that allows for a comparative TSS annotation by mapping RNAM-bM-^@M-^Sseq data of multiple strains into a common coordinate system derived from a whole-genome alignment. Considering the steadily increasing amount of RNAM-bM-^@M-^Sseq studies, our automated TSS annotation will not only facilitate transcriptome annotation for a wider range of pro- and eukaryotes but can also be adapted for the analysis among different growth or stress conditions. Our comparative dRNAM-bM-^@M-^Sseq analysis revealed conservation of most TSS, but also single-nucleotide-polymorphisms (SNP) in promoter regions, which lead to strain-specific transcriptional output. Furthermore, we identified strain-specific sRNA repertoires that could contribute to differential gene regulation among strains. In addition, we identified a novel minimal CRISPR-system in Campylobacter of the type-II CRISPR subtype, which relies on the host factor RNase III and a trans-encoded sRNA for maturation of crRNAs. This minimal system of Campylobacter, which seems active in only some strains, employs a unique maturation pathway, since the crRNAs are transcribed from individual promoters in the upstream repeats and thereby minimize the requirements for the maturation machinery. Overall, our study provides new insights into strain-specific transcriptome organization and sRNAs, and reveals genes that could modulate phenotypic variation among strains despite high conservation at the DNA level. Our dRNA-seq study of multiple C. jejuni strains represents the first comparative analysis of the primary transcriptomes of multiple strains and provides new insights into riboregulation in this bacterial pathogen.

			Action	DRS
	T12_MIC_1_cj.wiff	Wiff
	T12_MIC_1_cj.wiff.quant	Wiff
	T12_MIC_1_cj.wiff.scan	Wiff
	T12_MIC_1_cj.wiff2	Wiff
	T12_MIC_2_cj.wiff	Wiff

Dataset Information

Quantitative SWATH Proteomic Analysis of Campylobacter jejuni and Campylobacter coli Following Exposure to the Antimicrobial Ionophore PBT2

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