Analysis of a cloned sequence of Legionella pneumophila encoding a 38 kD metalloprotease possessing haemolytic and cytotoxic activities.
The DNA encoding the zinc metalloprotease of Legionella pneumophila Philadelphia 1 has been isolated and expressed in Escherichia coli. This protein, which is 38,000 Daltons in size, possesses immunological and biochemical properties identical to those previously described for the purified L. pneumophila protease. Periplasmic extracts of E. coli clones expressing the recombinant protease are also capable of causing the haemolysis of canine erythrocytes and the cytotoxic destruction of CHO cells. Using transposon mutagenesis, it was determined that a maximum of 1.2 kb of DNA encoded all three biological activities. Inactivation of proteolytic activity by transposon insertion occurred concomitantly with losses of the haemolytic and cytotoxic phenotypes. A putative regulatory sequence approximately 200-500 bp upstream of the gene's coding region was identified. A 4.0 kb fragment encoding these activities hybridized to the chromosomal DNA of the parent strain of L. pneumophila Philadelphia 1 as well as clinical isolates of L. pneumophila.
Quinn
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FD
Department of Microbiology and Immunology, Stanford University School of Medicine, California 94305.
Tompkins
L S
LS
D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D003360:Cosmids; D003603:Cytotoxins / Q000378:metabolism*; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D006461:Hemolysis; D007875:Legionella / Q000201:enzymology / Q000235:genetics*; D008666:Metalloendopeptidases / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping
D003603:Cytotoxins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D010447:Peptide Hydrolases; D008666:Metalloendopeptidases