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Published on: October 31, 2013
CcpA forms complexes with CodY and RpoA in Bacillus subtilis
Andrea Wünsche1, Elke Hammer, Maike Bartholomae
1Lehrstuhl für Mikrobiologie, Department Biologie, Friedrich-Alexander Universität-Erlangen-Nürnberg, Germany.
Bacillus subtilis catabolite control protein A (CcpA) interacts with RNA polymerase alpha-subunit (RpoA) and the regulator CodY, forming transcription initiation complexes. These interactions are crucial for gene regulation, especially under glucose availability.
Area of Science:
- Microbiology
- Molecular Biology
- Biochemistry
Background:
- Bacillus subtilis catabolite control protein A (CcpA) is a global regulator controlled by phosphoproteins and effectors.
- Mutations in CcpA affect target gene regulation, indicating additional protein interactions.
Purpose of the Study:
- To identify CcpA complexes involved in gene repression and activation.
- To elucidate the molecular mechanisms of CcpA-mediated transcriptional regulation.
Main Methods:
- In vivo crosslinking and mass spectrometry (MS) to identify protein complexes.
- Bacterial two-hybrid assays to determine CcpA binding sites.
- Surface plasmon resonance (SPR) to quantify binding affinities.
Main Results:
- CcpA interacts with RNA polymerase alpha-subunit (RpoA), HPr, and CodY.
- CcpA-RpoA complexes are present regardless of glucose, while CcpA-HPr and CcpA-CodY complexes form primarily with glucose.
- CcpA and CodY bind to both N-terminal (αNTD) and C-terminal (αCTD) domains of RpoA.
Conclusions:
- CcpA forms transcription initiation complexes with RpoA and CodY.
- These interactions are dynamic and influenced by carbon source availability.
- A model for the ackA promoter transcription initiation complex involving CcpA and CodY is proposed.
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