Reduced Glutathione Is Required for Pertussis Toxin Secretion by
            <i>Bordetella pertussis</i>

Stenson, Trevor H.; Patton, Angela K.; Weiss, Alison A.

doi:10.1128/iai.71.3.1316-1320.2003

Cited by 11 publications

(7 citation statements)

References 33 publications

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“…Since B. pertussis is an obligate aerobe, testing for direct oxygen oxidation is tech- nically not feasible. Cystine is a required growth factor for B. pertussis; thus, completely removing it from growth media is not possible (53,54). However, changing the levels of cystine did not perturb cytochrome c synthesis (not shown).…”

Section: And 5 [Example Of Results With Cb75])mentioning

confidence: 99%

Mutations in Cytochrome Assembly and Periplasmic Redox Pathways inBordetella pertussis

et al. 2005

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Transposon mutagenesis of Bordetella pertussis was used to discover mutations in the cytochrome c biogenesis pathway called system II. Using a tetramethyl-p-phenylenediamine cytochrome c oxidase screen, 27 oxidasenegative mutants were isolated and characterized. Nine mutants were still able to synthesize c-type cytochromes and possessed insertions in the genes for cytochrome c oxidase subunits (ctaC, -D, and -E), heme a biosynthesis (ctaB), assembly of cytochrome c oxidase (sco2), or ferrochelatase (hemZ). Eighteen mutants were unable to synthesize all c-type cytochromes. Seven of these had transposons in dipZ (dsbD), encoding the transmembrane thioreduction protein, and all seven mutants were corrected for cytochrome c assembly by exogenous dithiothreitol, which was consistent with the cytochrome c cysteinyl residues of the CXXCH motif requiring periplasmic reduction. The remaining 11 insertions were located in the ccsBA operon, suggesting that with the appropriate thiol-reducing environment, the CcsB and CcsA proteins comprise the entire system II biosynthetic pathway. Antiserum to CcsB was used to show that CcsB is absent in ccsA mutants, providing evidence for a stable CcsA-CcsB complex. No mutations were found in the genes necessary for disulfide bond formation (dsbA or dsbB). To examine whether the periplasmic disulfide bond pathway is required for cytochrome c biogenesis in B. pertussis, a targeted knockout was made in dsbB. The DsbB ؊ mutant makes holocytochromes c like the wild type does and secretes and assembles the active periplasmic alkaline phosphatase. A dipZ mutant is not corrected by a dsbB mutation. Alternative mechanisms to oxidize disulfides in B. pertussis are analyzed and discussed.

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Section: And 5 [Example Of Results With Cb75])mentioning

confidence: 99%

Mutations in Cytochrome Assembly and Periplasmic Redox Pathways inBordetella pertussis

et al. 2005

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“…The holotoxin contains 11 disulfide bonds: one in S1, two in S4 and S5, and three in S2 and S3. The periplasmic disulfide bond‐catalyzing proteins DsbA‐C and reduced glutathione disulfides are required for efficient PTX assembly [18,19]. Holotoxin assembly takes place in the periplasm even in the absence of the Ptl machinery, as shown by studies conducted both in vivo and in vitro [20–23].…”

Section: Biogenesis Of Ptxmentioning

confidence: 99%

The ins and outs of pertussis toxin

2011

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Pertussis toxin, produced and secreted by the whooping cough agent Bordetella pertussis, is one of the most complex soluble bacterial proteins. It is actively secreted through the B. pertussis cell envelope by the Ptl secretion system, a member of the widespread type IV secretion systems. The toxin is composed of five subunits (named S1 to S5 according to their decreasing molecular weights) arranged in an A-B structure. The A protomer is composed of the enzymatically active S1 subunit, which catalyzes ADP-ribosylation of the a subunit of trimeric G proteins, thereby disturbing the metabolic functions of the target cells, leading to a variety of biological activities. The B oligomer is composed of 1S2:1S3:2S4:1S5 and is responsible for binding of the toxin to the target cell receptors and for intracellular trafficking via receptor-mediated endocytosis and retrograde transport. The toxin is one of the most important virulence factors of B. pertussis and is a component of all current vaccines against whooping cough.

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“…2). It was reported that there is no difference in Prn production by Tohama I cultured in modified SS medium compared to that in a culture in modified SS medium containing DTT, a reducing agent (27). These results suggest that BvgAS-regulated virulence factors other than type III secreted proteins are not affected by reducing agents.…”

Section: Discussionmentioning

confidence: 73%

Transcriptional Downregulation of a Type III Secretion System under Reducing Conditions in Bordetella pertussis

et al. 2020

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Bordetella pertussis uses a type III secretion system (T3SS) to inject virulence proteins into host cells. Although the B. pertussis T3SS was presumed to be involved in host colonization, the efficient secretion of type III secreted proteins from B. pertussis has not been observed. To investigate the roles of type III secreted proteins during infection, we attempted to optimize culture conditions for the production and secretion of a type III secreted protein, BteA, in B. pertussis. We observed that B. pertussis efficiently secretes BteA in ascorbic acid-depleted (AsA-) medium. When L2 cells, a rat lung epithelial cell line, were infected with B. pertussis cultured in the AsA- medium, BteA-dependent cytotoxicity was observed. We also performed an immunofluorescence assay of L2 cells infected with B. pertussis. The clear fluorescence signals of Bsp22, a needle structure of T3SS, were detected on the bacterial surface of B. pertussis cultured in the AsA- medium. Since ascorbic acid is known as a reducing agent, we cultured B. pertussis in liquid medium containing other reducing agents such as 2-mercaptoethanol and dithioerythritol. Under these reducing conditions, the production of type III secreted proteins was repressed. These results suggest that in B. pertussis, the production and secretion of type III secreted proteins are downregulated under reducing conditions. IMPORTANCE The type III secretion system (T3SS) of Bordetella pertussis forms a needle-like structure that protrudes from the bacterial cell surface. B. pertussis uses T3SS to translocate virulence proteins called effectors into host cells. The culture conditions for effector production in B. pertussis have not been investigated. We attempted to optimize culture medium compositions for producing and secreting type III secreted proteins. We found that B. pertussis secretes type III secreted proteins in reducing agent-deprived liquid medium, and that BteA-secreting B. pertussis provokes cytotoxicity against cultured mammalian cells. These results suggest that redox signaling is involved in the regulation of B. pertussis T3SS.

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Reduced Glutathione Is Required for Pertussis Toxin Secretion by Bordetella pertussis

Cited by 11 publications

References 33 publications

Mutations in Cytochrome Assembly and Periplasmic Redox Pathways inBordetella pertussis

Mutations in Cytochrome Assembly and Periplasmic Redox Pathways inBordetella pertussis

The ins and outs of pertussis toxin

Transcriptional Downregulation of a Type III Secretion System under Reducing Conditions in Bordetella pertussis

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