Secretion Genes as Determinants of Bacillus anthracis Chain Length

• Published in: Journal of Bacteriology 2012-08, Vol.194 (15), p.3841-3850
• Main Authors: Nguyen-Mau, Sao-Mai, Oh, So-Young, Kern, Valerie J, Missiakas, Dominique M, Schneewind, Olaf
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphatases - genetics; Adenosine Triphosphatases - metabolism; Bacillus anthracis; Bacillus anthracis - cytology; Bacillus anthracis - genetics; Bacillus anthracis - growth & development; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Biological and medical sciences; Cells; chromosomes; Cytoplasm; Fundamental and applied biological sciences. Psychology; Gene Deletion; genes; Genes, Bacterial; Genetic Complementation Test; Membrane Glycoproteins - genetics; Membrane Glycoproteins - metabolism; Membrane Transport Proteins - genetics; Membrane Transport Proteins - metabolism; Membranes; Microbiology; Miscellaneous; Mutagenesis, Insertional; mutants; Mutation; peptidoglycans; Proteins; SEC Translocation Channels; SecA Proteins; secretion; Tissues
• ISSN: 0021-9193; 1098-5530; 1067-8832
• DOI: 10.1128/JB.00384-12

Bacillus anthracis grows in chains of rod-shaped cells, a trait that contributes to its escape from phagocytic clearance in host tissues. Using a genetic approach to search for determinants of B. anthracis chain length, we identified mutants with insertional lesions in secA2. All isolated secA2 mutants exhibited an exaggerated chain length, whereas the dimensions of individual cells were not changed. Complementation studies revealed that slaP (S-layer assembly protein), a gene immediately downstream of secA2 on the B. anthracis chromosome, is also a determinant of chain length. Both secA2 and slaP are required for the efficient secretion of Sap and EA1 (Eag), the two S-layer proteins of B. anthracis, but not for the secretion of S-layer-associated proteins or of other secreted products. S-layer assembly via secA2 and slaP contributes to the proper positioning of BslO, the S-layer-associated protein, and murein hydrolase, which cleaves septal peptidoglycan to separate chains of bacilli. SlaP was found to be both soluble in the bacterial cytoplasm and associated with the membrane. The purification of soluble SlaP from B. anthracis-cleared lysates did not reveal a specific ligand, and the membrane association of SlaP was not dependent on SecA2, Sap, or EA1. We propose that SecA2 and SlaP promote the efficient secretion of S-layer proteins by modifying the general secretory pathway of B. anthracis to transport large amounts of Sap and EA1.