Dataset Information

Insight into the interplay between thiol oxidoreductases of oxidative activity of Campylobacter jejuni

ABSTRACT: The bacterial proteins of the Dsb family catalyze the formation of disulfide bridges between cysteine residues that stabilize protein structures and ensure their proper functioning. Here we report the detailed analysis of the Dsb pathway of Campylobacter jejuni. The oxidizing Dsb system of this pathogen is unique because it consists of two monomeric DsbAs (DsbA1 and DsbA2) and one dimeric bifunctional protein (C8J_1298). Previously, we showed that DsbA1 and C8J_1298 are redundant. Here we unraveled the interaction between the two monomeric DsbAs by in vitro and in vivo experiments and by solving their structures and found that both monomeric DsbAs are dispensable proteins. Their structures confirmed that they are homologs of EcDsbL. The slight differences seen in the surface charge of the proteins do not affect the interaction with their redox partner. Comparative proteomics showed that several respiratory proteins, as well as periplasmic transport proteins, are targets of the Dsb system. Some of these, both donors and electron acceptors, are essential elements of C. jejuni respiratory process under oxygen-limiting conditions in the host intestine. The data presented provides detailed information on function of the C. jejuni Dsb system, identifying it as a potential target for novel antibacterial molecules.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Campylobacter Jejuni Subsp. Jejuni 81116

SUBMITTER: Agata Malinowska

LAB HEAD: Katarzyna Jagusztyn-Krynicka

PROVIDER: PXD029340 | Pride | 2022-02-22

REPOSITORIES: Pride

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Publications

Interplay between DsbA1, DsbA2 and C8J_1298 Periplasmic Oxidoreductases of <i>Campylobacter jejuni</i> and Their Impact on Bacterial Physiology and Pathogenesis.

Banaś Anna M AM Bocian-Ostrzycka Katarzyna M KM Dunin-Horkawicz Stanisław S Ludwiczak Jan J Wilk Piotr P Orlikowska Marta M Wyszyńska Agnieszka A Dąbrowska Maria M Plichta Maciej M Spodzieja Marta M Polańska Marta A MA Malinowska Agata A Jagusztyn-Krynicka Elżbieta Katarzyna EK

International journal of molecular sciences 20211215 24

The bacterial proteins of the Dsb family catalyze the formation of disulfide bridges between cysteine residues that stabilize protein structures and ensure their proper functioning. Here, we report the detailed analysis of the Dsb pathway of <i>Campylobacter jejuni</i>. The oxidizing Dsb system of this pathogen is unique because it consists of two monomeric DsbAs (DsbA1 and DsbA2) and one dimeric bifunctional protein (C8J_1298). Previously, we showed that DsbA1 and C8J_1298 are redundant. Here, ...[more]

PMID: 34948248

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Project description:C. jejuni, a spiral-shaped gram-negative bacterium, is a leading bacterial cause of human foodborne illness. Acute disease is associated with C. jejuni invasion of the intestinal epithelium. Further, maximal host cell invasion requires the secretion of proteins termed Campylobacter invasion antigens (Cia). As bile acids are known to alter the pathogenic behavior of other gastrointestinal pathogens, we hypothesized that the virulence potential of Campylobacter may be triggered by the bile acid deoxycholate (DOC). In support of this hypothesis, culturing C. jejuni with a physiologically relevant concentration of DOC significantly altered the kinetics of cell invasion as evidenced by gentamicin-protection assays. In contrast to C. jejuni harvested from Mueller-Hinton (MH) agar plates, C. jejuni harvested from MH agar plates supplemented with DOC demonstrated Cia secretion as judged by metabolic labeling experiments. DOC was also found to induce the expression of the ciaB gene as judged by B-galactosidase reporter assays and real-time RT-PCR. Microarray analysis revealed that DOC induced the expression of virulence genes (i.e., ciaB, cmeABC, dccR, and tlyA). In summary, we demonstrate that it is possible to enhance the pathogenic behavior of C. jejuni by modifying the culture conditions. These results provide a foundation to identify genes expressed by C. jejuni in response to in vivo-like culture conditions. Keywords: Stress response For the expression profiling arrays, an indirect comparison of gene expression was performed, where the expression profile of the C. jejuni F38011 cultured in the presence and absence of DOC was measured separately on different slides as described previously (26). Briefly, Cy5 labeled reference DNA from the C. jejuni F38011 strain was mixed with Cy3 labeled test cDNA (C. jejuni F38011 cultured in the presence or absence of DOC) and hybridized to the Campylobacter cDNA array (26) on separate slides. DNA microarrays were scanned using an Axon GenePix 4000B microarray laser scanner (Axon Instruments, Union City, CA) and the data for spot and background intensities were processed using the GenePix 4.0 software. To compensate for any effect of the amount of template and uneven Cy-dye incorporation, data normalization was performed as previously described (26). For the comparison of genes differentially expressed in the presence and absence of DOC,six hybridization measurements were generated per biological experiment (three technical replicate arrays and two replicate features per array).

Dataset Information

Insight into the interplay between thiol oxidoreductases of oxidative activity of Campylobacter jejuni

Publications

Interplay between DsbA1, DsbA2 and C8J_1298 Periplasmic Oxidoreductases of <i>Campylobacter jejuni</i> and Their Impact on Bacterial Physiology and Pathogenesis.

Similar Datasets

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