title: The Legionella pneumophila major secretory protein, a protease, is not required for intracellular growth or cell killing.
abstract: The Legionella pneumophila major secretory protein (Msp) is a Zn2+ metalloprotease whose function in pathogenesis is unknown. The structural gene for the Msp protease, mspA, was isolated from an L. pneumophila genomic library. In Escherichia coli which contain plasmids with the mspA gene, Msp protein and activity are found in the periplasmic space and the cytoplasm. Transposon mutagenesis with Tn9 of an mspA-containing plasmid in E. coli yielded mutants which no longer expressed protease activity and others with increased protease activity. These results suggested that mspA expression might be regulated. Msp was shown to be produced at a much higher level in L. pneumophila grown in rich compared to semidefined media. A Tn9 insertion which abolishes Msp expression was introduced into the L. pneumophila genome. This mspA::Tn9 L. pneumophila strain showed no detectable production of Msp by immunoblot analysis, and it had less than 0.1% of the protease activity found in the wild-type strain. This mutant was fully capable of growing within and killing human macrophages derived from the HL-60 cell line.
authors.affiliation: Department of Microbiology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
authors.forename: L
authors.initials: L
authors.lastname: Szeto
authors.forename: H A
authors.initials: HA
authors.lastname: Shuman
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D015139:Blotting, Southern; D002458:Cell Fractionation; D002470:Cell Survival; D002478:Cells, Cultured; D004251:DNA Transposable Elements; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D005798:Genes, Bacterial; D006801:Humans; D007875:Legionella / Q000201:enzymology* / Q000254:growth & development / Q000472:pathogenicity; D008264:Macrophages / Q000382:microbiology; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000235:genetics*; D009154:Mutation; D010641:Phenotype; D010957:Plasmids / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D008666:Metalloendopeptidases