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Journal of Bacteriology, February 2001, p. 1312-1319, Vol. 183, No. 4
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.4.1312-1319.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Disulfide Bond Formation in Secreton Component PulK Provides a Possible Explanation for the Role of DsbA in Pullulanase Secretion

Anthony P. Pugsley,* Nicolas Bayan, and Nathalie Sauvonnetdagger

Unité de Génétique Moléculaire, CNRS URA 1773-Institut Pasteur, 75724 Paris Cedex 15, France

Received 12 July 2000/Accepted 15 November 2000

When expressed in Escherichia coli, the 15 Klebsiella oxytoca pul genes that encode the so-called Pul secreton or type II secretion machinery promote pullulanase secretion and the assembly of one of the secreton components, PulG, into pili. Besides these pul genes, efficient pullulanase secretion also requires the host dsbA gene, encoding a periplasmic disulfide oxidoreductase, independently of disulfide bond formation in pullulanase itself. Two secreton components, the secretin pilot protein PulS and the minor pseudopilin PulK, were each shown to posses an intramolecular disulfide bond whose formation was catalyzed by DsbA. PulS was apparently destabilized by the absence of its disulfide bond, whereas PulK stability was not dramatically affected either by a dsbA mutation or by the removal of one of its cysteines. The pullulanase secretion defect in a dsbA mutant was rectified by overproduction of PulK, indicating reduced disulfide bond formation in PulK as the major cause of the secretion defect under the conditions tested (in which PulS is probably present in considerable excess of requirements). PulG pilus formation was independent of DsbA, probably because PulK is not needed for piliation.

* Corresponding author. Mailing address: Unité de Génétique moléculaire, Institut Pasteur, 25, rue du Dr. Roux, 75724 Paris CEDEX 15, France. Phone: 33/0-145688494. Fax: 33/0-145688960. E-mail: max{at}pasteur.fr.

dagger Present address: Microbial Pathogenesis Unit, Université Catholique de Louvain, B-1200 Brussels, Belgium.

Journal of Bacteriology, February 2001, p. 1312-1319, Vol. 183, No. 4
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.4.1312-1319.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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