1. MecA Protein Acts as a Negative Regulator of Genetic Competence in Streptococcus mutans
- Author
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Astrid Wahl, Zubelda A. Gomez, Xiao-Lin Tian, Yung-Hua Li, Tianlei Liu, Pascal Hols, and Gaofeng Dong
- Subjects
Transcription, Genetic ,Blotting, Western ,Biology ,Microbiology ,Streptococcus mutans ,Bacterial Proteins ,Genes, Reporter ,Transcription (biology) ,Sigma factor ,Two-Hybrid System Techniques ,Protein biosynthesis ,Post-translational regulation ,Luciferases ,Molecular Biology ,Gene ,Regulation of gene expression ,Genetics ,DNA Transformation Competence ,Gene Expression Regulation, Bacterial ,Articles ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,Artificial Gene Fusion ,Protein Biosynthesis ,Protein Processing, Post-Translational ,Gene Deletion - Abstract
Streptococcus mutans develops competence for genetic transformation through a complex network that receives inputs from at least two signaling peptides, competence-stimulating peptide (CSP) and sigX -inducing peptide (XIP). The key step of competence induction is the transcriptional activation of comX , which encodes an alternative sigma factor, SigX (σ X ), controlling the expression of late competence genes essential for DNA uptake and recombination. In this study, we provide evidence that MecA acts as a negative regulator in the posttranslational regulation of SigX in S. mutans . Using luxAB transcriptional reporter strains, we demonstrate that MecA represses the expression of late competence genes in S. mutans grown in a complex medium that is subpermissive for competence induction by CSP. The negative regulation of competence by MecA requires the presence of a functional SigX. Accordingly, inactivation of MecA results in a prolonged competence state of S. mutans under this condition. We have also found that the AAA+ protease ClpC displays a similar repressing effect on late competence genes, suggesting that both MecA and ClpC function coordinately to regulate competence in the same regulatory circuit in S. mutans . This suggestion is strongly supported by the results of bacterial two-hybrid assays, which demonstrate that MecA interacts with both SigX and ClpC, forming a ternary SigX-MecA-ClpC complex. Western blot analysis also confirms that inactivation of MecA or ClpC results in the intracellular accumulation of the SigX in S. mutans . Together, our data support the notion that MecA mediates the formation of a ternary SigX-MecA-ClpC complex that sequesters SigX and thereby negatively regulates genetic competence in S. mutans .
- Published
- 2013
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