Identification, cloning, and characterization of the lma Operon, whose gene products are unique to Listeria monocytogenes

The lmaA gene of Listeria monocytogenes encodes a protein capable of inducing delayed-type hypersensitivity reactions in ii, monocytogenes-immune mice (S. Gohmann, M. Leimeister-Wachter, E. Schiltz, W. Goebel, and T. Chakraborty, M. Microbiol, 4:1091-1099, 1990), Here we show that it is the last gene of the lma operon, which now comprises four genes, lmaDCBA. Maxicell analysis of peptides encoded by the Ima operon identified four polypeptides of 16.7, 16.4, 14.9, and 21 kDa which correspond to the gene products encoded by the lmaD, -C, -B, and -A genes, respectively, Northern blot analysis of the lma operon showed that lmaA is expressed by two transcripts: the longer lmaDCBA transcript of 2,100 nucleotides, which was observed at growth temperatures of 37 and 20 degrees C, and a shorter transcript consisting of lmaBA, which is detected only at low temperatures (20 degrees C), Two promoters, one preceding the lmaD gene and another located upstream of the lmaB gene, were detected, An extended stem-loop structure resembling box elements found in other gram-positive pathogens was also present in the lmaC-lmaB intergenic region, By immunoblot analysis, we found that although LmaA was produced at both temperatures (20 and 37 degrees C), it was secreted into culture supernatants only at 20 degrees C, However, LmaA lacks a bona fide signal peptide sequence and could, like flagellin, be secreted by a type III transport system, DNA hybridization studies indicate that the lma operon is species specific and restricted to pathogenic strains of L. monocytogenes.