A general strategy for identification of S-layer genes in the Bacillus cereus group: molecular characterization of such a gene in Bacillus thuringiensis subsp. galleriae NRRL 4045

• Published in: Microbiology (Society for General Microbiology) Vol. 147; no. 5; pp. 1343 - 1351
• Main Authors: Mesnage, Stephane, Haustant, Michel, Fouet, Agnes
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01.05.2001; Society for General Microbiology
• Subjects: Amino Acid Motifs; Amino Acid Sequence; Bacillus cereus - genetics; Bacillus cereus - metabolism; Bacillus thuringiensis - genetics; Bacillus thuringiensis - metabolism; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Biological and medical sciences; Cell Wall - metabolism; Fundamental and applied biological sciences. Psychology; Genes, Bacterial - genetics; Genetics; Microbiology; Molecular Sequence Data; Mutagenesis, Insertional; Peptidoglycan - metabolism; Protein Structure, Tertiary; Sequence Alignment; Sequence Homology, Amino Acid; Membrane protein; Nucleotide sequence; Bacillus thuringiensis; Binding site; Bacillaceae; External membrane; Cell surface; Cell wall; Antigen; Bacillus cereus; Bacillales; Structural unit; Bacteria; S Layer; Aminoacid sequence
• ISSN: 1350-0872; 1465-2080
• DOI: 10.1099/00221287-147-5-1343

Toxines et Pathogénie Bactériennes (URA 2172, CNRS), Institut Pasteur, 28 rue du Dr Roux, 75724, Paris cédex 15, France 1 Author for correspondence: Stéphane Mesnage. Tel: +44 1603 450514. Fax: +44 1603 450025. e-mail: stephane.mesnage{at}bbsrc.ac.uk Despite its possible role in virulence, there has been little molecular characterization of members of the S-layer protein family of the Bacillus cereus group. It is hypothesized that the components of the S-layers are likely to display similar anchoring structures in Bacillus thuringiensis and Bacillus anthracis . Based on inferred sequence similarities, a DNA fragment encoding the cell-wall-anchoring domain of an S-layer component of Bac. thuringiensis subsp. galleriae NRRL 4045 was isolated. The complete gene was identified and sequenced. An ORF of 2463 nt was identified, which was predicted to encode a protein of 821 aa, SlpA. The mature SlpA protein (792 residues) carries three S-layer homology (SLH) motifs towards its amino terminus, each about 50 aa long. These motifs were sufficient to bind Bac. thuringiensis and Bac. anthracis cell walls in vitro by interacting with peptidoglycan-associated polymers, confirming a common wall-anchoring mechanism. The slpA null-allele mutant was constructed and shown to possess no other abundant surface protein. It is proposed that the method described in this paper could be applied to the identification and deletion of any Bac. cereus S-layer gene and is of great value for the molecular and functional characterization of these genes. Keywords: SLH motifs, cell-wall anchoring, surface antigens, Gram-positive The GenBank accession number for the slpA sequence is AJ249446 . a Present address: John Innes Centre, Department of Cell and Developmental Biology, Colney Lane, Norwich NR4 7UH, UK.