title: Are the PE-PGRS proteins of Mycobacterium tuberculosis variable surface antigens?
abstract: Mycobacterium tuberculosis H37Rv contains 67 PE-PGRS genes, with multiple tandem repetitive sequences, encoding closely related proteins that are exceptionally rich in glycine and alanine. As no functional information was available, 10 of these genes were selected and shown to be expressed in vitro by reverse transcription-polymerase chain reaction (RT-PCR). Antibodies against five PE-PGRS proteins, raised in mice by DNA vaccination, detected single proteins when the same plasmid constructs used for immunization were expressed in epithelial cells or in reticulocyte extracts, confirming that the PE-PGRS proteins are antigenic. As expected from the conserved repetitive structure, the antibodies cross-reacted with more than one PE-PGRS protein, suggesting that different proteins share common epitopes. PE-PGRS proteins were detected by West-ern blotting in five different mycobacterial species (M. tuberculosis, M. bovis BCG, M. smegmatis, M. marinum and M. gordonae) and 11 clinical isolates of M. tuberculosis. Whole-genome comparisons of M. tuberculosis predicted allelic diversity in the PE-PGRS family, and this was confirmed by immunoblot studies as size variants were detected in clinical strains. Subcellular fractionation studies and immunoelectron microscopy localized many PE-PGRS proteins in the cell wall and cell membrane of M. tuberculosis. The data suggest that some PE-PGRS proteins are variable surface antigens.
authors.affiliation: Unité de Génétique Moléculaire Bactérienne, Institut Pasteur, 28 rue du Docteur Roux, 75724 Paris Cedex 15, France.
authors.forename: Sayera
authors.initials: S
authors.lastname: Banu
authors.forename: Nadine
authors.initials: N
authors.lastname: Honoré
authors.forename: Brigitte
authors.initials: B
authors.lastname: Saint-Joanis
authors.forename: Dana
authors.initials: D
authors.lastname: Philpott
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Prévost
authors.forename: Stewart T
authors.initials: ST
authors.lastname: Cole
mesh_terms: D000942:Antigens, Bacterial / Q000032:analysis*; D001426:Bacterial Proteins / Q000235:genetics* / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D007118:Immunoassay; D008565:Membrane Proteins / Q000235:genetics* / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000302:isolation & purification; D012156:Reticulocytes / Q000382:microbiology; D014162:Transfection
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D008565:Membrane Proteins; C440642:PE-PGRS protein, Mycobacterium