Enhanced expression of lmb gene encoding laminin-binding protein in Streptococcus agalactiae strains harboring IS1548 in scpB-lmb intergenic region.
Group B streptococcus (GBS) is the main cause of neonatal sepsis and meningitis. Bacterial surface proteins play a major role in GBS binding to and invasion of different host surfaces. The scpB and lmb genes, coding for fibronectin-binding and laminin-binding surface proteins, are present in almost all human GBS isolates. The scpB-lmb intergenic region is a hot spot for integration of two mobile genetic elements (MGEs): the insertion element IS1548 or the group II intron GBSi1. We studied the structure of scpB-lmb intergenic region in 111 GBS isolates belonging to the intraspecies major clonal complexes (CCs). IS1548 was mostly found (72.2%) in CC19 serotype III strains recovered more specifically (92.3%) from neonatal meningitis. GBSi1 was principally found (70.6%) in CC17 strains, mostly (94.4%) of serotype III, but also (15.7%) in CC19 strains, mostly (87.5%) of serotype II. No MGE was found in most strains of the other CCs (76.0%), notably CC23, CC10 and CC1. Twenty-six strains representing these three genetic configurations were selected to investigate the transcription and expression levels of scpB and lmb genes. Quantitative RT-PCR showed that lmb transcripts were 5.0- to 9.6-fold higher in the group of strains with IS1548 than in the other two groups of strains (P<0.001). Accordingly, the binding ability to laminin was 3.8- to 6.6-fold higher in these strains (P< or =0.001). Moreover, Lmb amount expressed on the cell surface was 2.4- to 2.7-fold greater in these strains (P<0.001). By contrast, scpB transcript levels and fibronectin binding ability were similar in the three groups of strains. Deletion of the IS1548 sequence between scpB and lmb genes in a CC19 serotype III GBS strain substantially reduced the transcription of lmb gene (13.5-fold), the binding ability to laminin (6.2-fold), and the expression of Lmb protein (5.0-fold). These data highlight the importance of MGEs in bacterial virulence and demonstrate the up-regulation of lmb gene by IS1548; the increased lmb gene expression observed in CC19 serotype III strains with IS1548 may play a role in their ability to cause neonatal meningitis and endocarditis.
Equipe d'Accueil 3854 Bactéries et Risque Materno-Foetal, Institut Fédératif de Recherche 136 Agents Transmissibles et Infectiologie, UFR Médecine, Université François Rabelais de Tours, Tours, France.
Rim
R
Al Safadi
Souheila
S
Amor
Geneviève
G
Hery-Arnaud
Barbara
B
Spellerberg
Philippe
P
Lanotte
Laurent
L
Mereghetti
François
F
Gannier
Roland
R
Quentin
Agnès
A
Rosenau
D018829:Adhesins, Bacterial / Q000235:genetics*; D016667:Bacterial Capsules / Q000145:classification; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D015373:Bacterial Typing Techniques; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D021901:DNA, Intergenic / Q000235:genetics*; D010450:Endopeptidases / Q000235:genetics*; D005353:Fibronectins / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial / Q000235:genetics*; D006801:Humans; D055767:Immobilized Proteins / Q000378:metabolism; D020071:Interspersed Repetitive Sequences / Q000235:genetics; D007797:Laminin / Q000378:metabolism; D016254:Mutagenesis, Insertional / Q000235:genetics*; D010802:Phylogeny; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D013292:Streptococcus agalactiae / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D021901:DNA, Intergenic; D005353:Fibronectins; D055767:Immobilized Proteins; D007797:Laminin; D012333:RNA, Messenger; D010450:Endopeptidases; C053088:C5a peptidase