Pseudomonas aeruginosa alkaline protease: evidence for secretion genes and study of secretion mechanism.
A 6.5-kb DNA fragment carrying the functions required for specific secretion of the extracellular alkaline protease produced by Pseudomonas aeruginosa was cloned. The whole 6.5-kb DNA fragment was transcribed in one direction and probably carried three genes involved in secretion. The expression in trans of these genes, together with the apr gene, in Escherichia coli allowed synthesis and secretion of the alkaline protease, which was extensively investigated by performing pulse-chase experiments under various conditions. We demonstrated the absence of a precursor form, as well as the independence of alkaline protease translocation from SecA. The absence of secretion genes impaired alkaline protease secretion; the protein then remained intracellular and was partially degraded.
Guzzo
J
J
Laboratoire de Chimie Bactérienne, C.N.R.S., Marseille, France.
Pages
J M
JM
Duong
F
F
Lazdunski
A
A
Murgier
M
M
D000251:Adenosine Triphosphatases / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D015153:Blotting, Western; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D026901:Membrane Transport Proteins; D010957:Plasmids; D011233:Precipitin Tests; D011498:Protein Precursors / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics; D015183:Restriction Mapping; D000069816:SEC Translocation Channels; D000081416:SecA Proteins; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; D011498:Protein Precursors; D000069816:SEC Translocation Channels; D012697:Serine Endopeptidases; C032923:microbial serine proteinases; D000251:Adenosine Triphosphatases; D000081416:SecA Proteins