Molecular cloning, sequencing, and identification of a metalloprotease gene from Listeria monocytogenes that is species specific and physically linked to the listeriolysin gene.
The entire nucleotide sequence of an open reading frame located immediately downstream of the listeriolysin gene from a virulent Listeria monocytogenes serotype 1/2a strain was determined. The product of the open reading frame was 510 amino acids with a predicted molecular weight of 57,400. The deduced amino acid sequence of this open reading frame is highly similar to that of a family of secreted metalloproteases produced by various members of the genus Bacillus, of which thermolysin is the prototype. Immunoblots performed with specific antisera raised against thermolysin from Bacillus stearothermophilus allowed the detection of a 60-kDa polypeptide, corresponding to the pro-form of the protease, in culture supernatants of L. monocytogenes strains. In maxicell experiments, Escherichia coli recombinants harboring this open reading frame also specifically directed production of a 60-kDa protein. Protease activity was low to undetectable in both Listeria strains and E. coli recombinants. This is due to lack of processing of the inactive pro-form of the protease to its mature active form in both species. We have designated this gene mpl for metalloprotease of L. monocytogenes. The gene was present only in pathogenic L. monocytogenes strains, in which it was physically linked to the listeriolysin gene.
Domann
E
E
Institut für Genetik und Mikrobiologie, Universität Würzburg, Federal Republic of Germany.
Leimeister-Wächter
M
M
Goebel
W
W
Chakraborty
T
T
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins; D001483:Base Sequence; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D006360:Heat-Shock Proteins / Q000235:genetics*; D006460:Hemolysin Proteins; D008089:Listeria monocytogenes / Q000201:enzymology / Q000235:genetics*; D008666:Metalloendopeptidases / Q000032:analysis / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D013045:Species Specificity
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; D008666:Metalloendopeptidases; C038705:hlyA protein, Listeria monocytogenes