Table 2. Chromosomal Genes Coding for Toxins in B. Cereus ATCC 14579 and B. Anthracis A2012

Table 2. Chromosomal genes coding for toxins in B. cereus ATCC 14579 and B. anthracis A2012. CDSs in Bce CDSs in Ban Function Reference BC3103*, BC3102*, Haemolytic enterotoxin 1 BC3102* BL BC1809*, BC1810*, BA_2389*, BA_2390*, Non-haemolytic 2 BC0560† BA_1129 enterotoxin Nhe BC2081 Enterotoxin T, BceT‡ 3 BC1953 BA_2454 Enterotoxin FM1 4 BC1110* Cytotoxin K 5 BA_4362 (upstream Phosphatidylinositol- BC3761* 6 sequence not known) specific phospholipase C

BC0670* Phosphatidylcholine- 7 specific phospholipase C BC0671* BA_1265* Sphingomyelinase 7 BC5101* BA_3854 Cereolysin O BC3523 BA_3024 * Hemolysin II 8 BC2196 Hemolysin III 9

* Potential PlcR-binding site was identified upstream of the gene(s). † Putative binding component of the non-hemolytic enterotoxin, BC0560, is not clustered with the nhe operon

(BC1809, BC1810). A PlcR-binding site was found upstream of both the nhe operon and BC0560.

‡ Diarrheal toxin BceT, which is commonly used for PCR-based detection of potentially enterotoxigenic Bce in foods, appears to be a part of a larger protein conserved in other food- borne pathogens, such as Staphylococci and Listeria. The toxins unlikely to be produced by the strain ATCC 14579 are the emetic toxin cereulide10 and the ADP-ribosylating toxin11. No CDS for the ADP-ribosylating toxin was found in the Bce genome. The genes encoding enzyme(s) for biosynthesis of the cyclic dodecadepsipeptide cereulide have not been identified so far. A cluster of four genes with similarity to peptide synthetase subunits was found in the genome, but none of them has the racemase domain, so the cluster is unlikely to code for biosynthesis of cereulide, which contains D-O-Leu and D-Ala. 1. Økstad, O. A. et al. Sequence analysis of three Bacillus cereus loci carrying PlcR-regulated

genes encoding degradative enzymes and enterotoxin. Microbiology 145, 3129-3138 (1999)

2. Lund, T. & Granum, P. E. Comparison of biological effect of the two different enterotoxin

complexes isolated from three different strains of Bacillus cereus. Microbiology 143, 3329-

3336 (1997)

3. Agata, N., Ohta, M., Arakawa, Y. & Mori, M. The bceT gene of Bacillus cereus encodes an

enterotoxic protein. Microbiology 141, 983-988 (1995)

4. Asano, S. I., Nukumizu, Y., Bando, H., Iizuka, T. & Yamamoto, T. Cloning of novel

enterotoxin genes from Bacillus cereus and Bacillus thuringiensis. Appl. Environ. Microbiol.

63, 1054-1057 (1997)

5. Lund, T., De Buyser, M. L. & Granum, P. E. A new cytotoxin from Bacillus cereus that may

cause necrotic enteritis. Mol. Microbiol. 38, 254-261 (2000)

6. Kuppe, A., Evans, L. M., McMillen, D. A. & Griffith, O. H. Phosphatidylinositol-specific

phospholipase C of Bacillus cereus: cloning, sequencing, and relationship to other

phospholipases. J. Bacteriol. 171, 6077-6083 (1989)

7. Gilmore, M. S., Cruz-Rodz, A. L., Leimeister-Waechter, M., Kreft, J. & Goebel, W. A Bacillus

cereus cytolytic determinant, cereolysin AB, which comprises the phospholipase C and

sphingomyelinase genes: nucleotide sequence and genetic linkage. J. Bacteriol. 171, 744-

753 (1989)

8. Baida, G., Budarina, Z. I., Kuzmin, N. P. & Solonin, A. S. Complete nucleotide sequence and

molecular characterization of hemolysin II gene from Bacillus cereus. FEMS Microbiol. Lett.

180, 7-14 (1999)

9. Baida, G. E. & Kuzmin, N. P. Cloning and primary stucture of a new hemolysin gene from

Bacillus cereus. Biochim. Biophys. Acta 1264, 151-154 (1995)

10. Agata, N. et al. A novel dodecadepsipeptide, cereulide, isolated from Bacillus cereus causes

vacuole formation in HEp-2 cells. FEMS Microbiol. Lett. 121, 31-34 (1994) 11. Just, I. et al. Rho-ADP-ribosylating exoenzyme from Bacillus cereus. Purification,

characterization, and identification of the NAD-binding site. Biochemistry 34, 334-340 (1995)