Enhanced expression of lmb gene encoding laminin-binding protein in Streptococcus agalactiae strains harboring IS1548 in scpB-lmb intergenic region

PLoS One. 2010 May 24;5(5):e10794. doi: 10.1371/journal.pone.0010794.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adhesins, Bacterial / genetics*
  • Bacterial Capsules / classification
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Bacterial Typing Techniques
  • Blotting, Western
  • Cell Membrane / metabolism
  • DNA, Intergenic / genetics*
  • Endopeptidases / genetics*
  • Fibronectins / metabolism
  • Gene Expression Regulation, Bacterial*
  • Genes, Bacterial / genetics*
  • Humans
  • Immobilized Proteins / metabolism
  • Interspersed Repetitive Sequences / genetics
  • Laminin / metabolism
  • Mutagenesis, Insertional / genetics*
  • Phylogeny
  • Protein Binding
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Sequence Analysis, DNA
  • Sequence Deletion
  • Streptococcus agalactiae / classification
  • Streptococcus agalactiae / genetics*
  • Streptococcus agalactiae / isolation & purification

Substances

  • Adhesins, Bacterial
  • Bacterial Proteins
  • DNA, Intergenic
  • Fibronectins
  • Immobilized Proteins
  • Laminin
  • RNA, Messenger
  • Endopeptidases
  • C5a peptidase