Disulfide bond formation in secreton component PulK provides a possible explanation for the role of DsbA in pullulanase secretion.
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.
Pugsley
A P
AP
Unité de Génétique Moléculaire, CNRS URA 1773-Institut Pasteur, 75724 Paris Cedex 15, France. max@pasteur.fr
Bayan
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N
Sauvonnet
N
N
D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D004220:Disulfides / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D036781:Fimbriae Proteins; D006026:Glycoside Hydrolases / Q000378:metabolism*; D007709:Klebsiella / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D019704:Protein Disulfide-Isomerases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004220:Disulfides; D008565:Membrane Proteins; C422472:PulK protein, Klebsiella oxytoca; C422473:PulS protein, Klebsiella oxytoca; D011994:Recombinant Proteins; D036781:Fimbriae Proteins; D006026:Glycoside Hydrolases; C100181:pullulanase; D019704:Protein Disulfide-Isomerases