PulO, a component of the pullulanase secretion pathway of Klebsiella oxytoca, correctly and efficiently processes gonococcal type IV prepilin in Escherichia coli.
The PulO protein required for extracellular secretion of pullulanase by Klebsiella oxytoca is known to be highly homologous to two type IV prepilin peptidases, namely XcpA(PilD) (Pseudomonas aeruginosa) and TcpJ (Vibrio cholerae). The predicted prepilin peptidase activity of PulO was confirmed by showing that it could correctly process the product of the cloned pilE.1 type IV pilin structural gene from Neisseria gonorrhoeae in Escherichia coli. The P. aeruginosa prepilin peptidase and another putative prepilin peptidase, ComC from Bacillus subtilis, also processed prePilE. Subcellular fractionation showed that the pilE gene product that had been processed by PulO remained associated with the cytoplasmic membrane, as did the unprocessed precursor. PulO was also shown to process three of the four prePilE-PhoA hybrids tested. Southern hybridization experiments suggest that a pulO homologue is present in the N. gonorrhoeae chromosome.
Unité des Neisseria, Institut Pasteur, Paris, France.
B
B
Dupuy
M K
MK
Taha
O
O
Possot
C
C
Marchal
A P
AP
Pugsley
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics*; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000201:enzymology / Q000378:metabolism*; D005798:Genes, Bacterial / Q000502:physiology*; D006026:Glycoside Hydrolases / Q000378:metabolism; D007700:Kinetics; D007709:Klebsiella / Q000235:genetics; D008954:Models, Biological; D009344:Neisseria gonorrhoeae / Q000235:genetics; D009693:Nucleic Acid Hybridization; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D011498:Protein Precursors; D011994:Recombinant Proteins; D036781:Fimbriae Proteins; D006026:Glycoside Hydrolases; C100181:pullulanase; D010450:Endopeptidases; C074273:type IV prepilin peptidase