Multiple roles of the pilus biogenesis protein pilD: involvement of pilD in excretion of enzymes from Pseudomonas aeruginosa.
In Pseudomonas aeruginosa, the genes pilB, pilC, and pilD encode proteins necessary for posttranslational modification and assembly of pilin monomers into pilus organelles (D. Nunn, S. Bergman, and S. Lory, J. Bacteriol. 172:2911-2919, 1990). We show that PilD, encoding a putative pilin-specific leader peptidase, also controls export of alkaline phosphatase, phospholipase C, elastase, and exotoxin A. pilD mutants accumulate these proteins in the periplasmic space, while secretion of periplasmic and outer membrane proteins appears to be normal. The periplasmic form of exotoxin A was fully mature in size, contained all cysteines in disulfide bonds, and was toxic in a tissue culture cytotoxicity assay, suggesting that in pilD mutants, exotoxin A was folded into its native conformation. The function of the other two accessory proteins, PilB and PilC, appears to be restricted to pilus biogenesis, and strains carrying mutations in their respective genes do not show an export defect. These studies show that in addition to cleaving the leader sequence from prepilin, PilD has an additional role in secretion of proteins that are released from P. aeruginosa into the surrounding media. PilD most likely functions as a protease that is involved in processing and assembly of one or more components of the membrane machinery necessary for the later stages of protein extracellular localization.
Strom
M S
MS
Department of Microbiology, School of Medicine, University of Washington, Seattle 98195.
Nunn
D
D
Lory
S
S
D036002:ADP Ribose Transferases; D000469:Alkaline Phosphatase / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D001427:Bacterial Toxins; D010450:Endopeptidases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D005098:Exotoxins / Q000378:metabolism; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000378:metabolism*; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D008565:Membrane Proteins; D009154:Mutation; D010196:Pancreatic Elastase / Q000378:metabolism; D011487:Protein Conformation; D011550:Pseudomonas aeruginosa / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D012697:Serine Endopeptidases; D010738:Type C Phospholipases / Q000378:metabolism; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A
D036002:ADP Ribose Transferases; D000469:Alkaline Phosphatase; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D010450:Endopeptidases; D005098:Exotoxins; D036781:Fimbriae Proteins; D008565:Membrane Proteins; D010196:Pancreatic Elastase; D012697:Serine Endopeptidases; D010738:Type C Phospholipases; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A; C067127:prepilin peptidase protein, Bacteria; C026515:type I signal peptidase