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Enrichment and Detection of Clostridium perfringens Toxinotypes in Retail Food Samples
Published on: October 18, 2019
A novel toxin regulator, the CPE1446-CPE1447 protein heteromeric complex, controls toxin genes in Clostridium
1Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba City, Ibaraki 305-8572, Japan. obana.nozomu.gb@u.tsukuba.ac.jp
Abstract:
Clostridium perfringens is a Gram-positive anaerobic spore-forming bacterium that is widespread in environmental soil and sewage, as well as in animal intestines. It is also a causative agent of diseases in humans and other animals, and it produces numerous extracellular enzymes and toxins. Although these toxins have been characterized in detail, regulators of toxin genes are less well understood. The present study identified CPE1447 and CPE1446 as novel regulators of toxin gene expression. CPE1447 and CPE1446 are cotranscribed as an operon, and the encoded proteins have a helix-turn-helix (HTH) motif at the N termini of their amino acid sequences, suggesting that CPE1447 and CPE1446 control the target genes as transcriptional regulators. The expression of several genes encoding toxins was changed in both a CPE1446 mutant and a CPE1447-CPE1446 deletion mutant. Complementation of CPE1446 and CPE1447 revealed that CPE1447 and CPE1446 coordinately regulate their target genes. CPE1447 protein was coprecipitated with His-tagged CPE1446 protein, indicating that the CPE1447 and CPE1446 proteins form a stable complex in C. perfringens under their native conditions. Although the small RNA that regulates several genes under the VirR/VirS two-component system (VR-RNA) positively affected CPE1447-CPE1446 mRNA expression, it did not control expression of the CPE1447-CPE1446 regulon, demonstrating that CPE1447 and CPE1446 regulate a different set of toxin genes from the VirR/VirS-VR-RNA cascade.
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