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Phosphatidylcholine-specific phospholipase C and sphingomyelinase activities in bacteria of the Bacillus cereus group.
Infect Immun 2003; 71(11):6591-606II

Abstract

Bacillus anthracis is nonhemolytic, even though it is closely related to the highly hemolytic Bacillus cereus. Hemolysis by B. cereus results largely from the action of phosphatidylcholine-specific phospholipase C (PC-PLC) and sphingomyelinase (SPH), encoded by the plc and sph genes, respectively. In B. cereus, these genes are organized in an operon regulated by the global regulator PlcR. B. anthracis contains a highly similar cereolysin operon, but it is transcriptionally silent because the B. anthracis PlcR is truncated at the C terminus. Here we report the cloning, expression, purification, and enzymatic characterization of PC-PLC and SPH from B. cereus and B. anthracis. We also investigated the effects of expressing PlcR on the expression of plc and sph. In B. cereus, PlcR was found to be a positive regulator of plc but a negative regulator of sph. Replacement of the B. cereus plcR gene by its truncated orthologue from B. anthracis eliminated the activities of both PC-PLC and SPH, whereas introduction into B. anthracis of the B. cereus plcR gene with its own promoter did not activate cereolysin expression. Hemolytic activity was detected in B. anthracis strains containing the B. cereus plcR gene on a multicopy plasmid under control of the strong B. anthracis protective antigen gene promoter or in a strain carrying a multicopy plasmid containing the entire B. cereus plc-sph operon. Slight hemolysis and PC-PLC activation were found when PlcR-producing B. anthracis strains were grown under anaerobic-plus-CO(2) or especially under aerobic-plus-CO(2) conditions. Unmodified parental B. anthracis strains did not demonstrate obvious hemolysis under the same conditions.

Authors+Show Affiliations

National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892-4350, USA.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

14573681

Citation

Pomerantsev, A P., et al. "Phosphatidylcholine-specific Phospholipase C and Sphingomyelinase Activities in Bacteria of the Bacillus Cereus Group." Infection and Immunity, vol. 71, no. 11, 2003, pp. 6591-606.
Pomerantsev AP, Kalnin KV, Osorio M, et al. Phosphatidylcholine-specific phospholipase C and sphingomyelinase activities in bacteria of the Bacillus cereus group. Infect Immun. 2003;71(11):6591-606.
Pomerantsev, A. P., Kalnin, K. V., Osorio, M., & Leppla, S. H. (2003). Phosphatidylcholine-specific phospholipase C and sphingomyelinase activities in bacteria of the Bacillus cereus group. Infection and Immunity, 71(11), pp. 6591-606.
Pomerantsev AP, et al. Phosphatidylcholine-specific Phospholipase C and Sphingomyelinase Activities in Bacteria of the Bacillus Cereus Group. Infect Immun. 2003;71(11):6591-606. PubMed PMID: 14573681.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Phosphatidylcholine-specific phospholipase C and sphingomyelinase activities in bacteria of the Bacillus cereus group. AU - Pomerantsev,A P, AU - Kalnin,K V, AU - Osorio,M, AU - Leppla,S H, PY - 2003/10/24/pubmed PY - 2003/12/3/medline PY - 2003/10/24/entrez SP - 6591 EP - 606 JF - Infection and immunity JO - Infect. Immun. VL - 71 IS - 11 N2 - Bacillus anthracis is nonhemolytic, even though it is closely related to the highly hemolytic Bacillus cereus. Hemolysis by B. cereus results largely from the action of phosphatidylcholine-specific phospholipase C (PC-PLC) and sphingomyelinase (SPH), encoded by the plc and sph genes, respectively. In B. cereus, these genes are organized in an operon regulated by the global regulator PlcR. B. anthracis contains a highly similar cereolysin operon, but it is transcriptionally silent because the B. anthracis PlcR is truncated at the C terminus. Here we report the cloning, expression, purification, and enzymatic characterization of PC-PLC and SPH from B. cereus and B. anthracis. We also investigated the effects of expressing PlcR on the expression of plc and sph. In B. cereus, PlcR was found to be a positive regulator of plc but a negative regulator of sph. Replacement of the B. cereus plcR gene by its truncated orthologue from B. anthracis eliminated the activities of both PC-PLC and SPH, whereas introduction into B. anthracis of the B. cereus plcR gene with its own promoter did not activate cereolysin expression. Hemolytic activity was detected in B. anthracis strains containing the B. cereus plcR gene on a multicopy plasmid under control of the strong B. anthracis protective antigen gene promoter or in a strain carrying a multicopy plasmid containing the entire B. cereus plc-sph operon. Slight hemolysis and PC-PLC activation were found when PlcR-producing B. anthracis strains were grown under anaerobic-plus-CO(2) or especially under aerobic-plus-CO(2) conditions. Unmodified parental B. anthracis strains did not demonstrate obvious hemolysis under the same conditions. SN - 0019-9567 UR - https://www.unboundmedicine.com/medline/citation/14573681/Phosphatidylcholine_specific_phospholipase_C_and_sphingomyelinase_activities_in_bacteria_of_the_Bacillus_cereus_group_ L2 - http://iai.asm.org/cgi/pmidlookup?view=long&pmid=14573681 DB - PRIME DP - Unbound Medicine ER -