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Research Article
Yongbin Xu
Dalian Minzu University
Corresponding Author
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Catabolite control protein C (CcpC) belongs to the LysR-type transcriptional regulator (LTTR) family that regulates the transcription of genes encoding the tricarboxylic acid branch enzymes of the TCA cycle by responding to a pathway-specific metabolite, citrate. The biological function of CcpC has been characterized several times, but the structural basis for the molecular function of CcpC remains elusive. Here, we report characterization of a full-length CcpC from Bacillus amyloliquefaciens (BaCcpC-FL) and a crystal structure of the C-terminal inducer-binding domain (IBD) complexed with citrate. BaCcpC required both dyad symmetric regions I and II to recognize the citB promoter, and the presence of citrate reduced citB promoter binding. The crystal structure of CcpC-IBD shows two subdomains, IBD-I and IBD-II, and a citrate molecule buried between them. Ile100, two arginines (Arg147 and Arg260), and three serines (Ser129, Ser189, and Ser191) have strong hydrogen-bond interactions with citrate molecules. A structural comparison of BaCcpC-IBD with its homologues shows that they share the same tail-to-tail dimer alignment, but the dimeric interface and the rotation between these molecules exhibit significant differences. In addition, citrate can convert large BaCcpC-FL oligomers to monomers in solution. Taken together, our results provide a framework for understanding the mechanism underlying the functional divergence of the CcpC protein.
Keywords
Catabolite control protein C (CcpC), Bacillus amyloliquefaciens (BaCcpC-FL)
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This preprint is under consideration at Scientific Reports. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research Article
Yongbin Xu
Dalian Minzu University
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 1
Posted 12 Mar, 2021
No community comments so far
Reviewers invited
Invitations sent on 10 Mar, 2021
Editor assigned
On 10 Mar, 2021
Editor invited
On 04 Mar, 2021
Submission checks complete
On 04 Mar, 2021
First submitted
On 01 Mar, 2021
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Catabolite control protein C (CcpC) belongs to the LysR-type transcriptional regulator (LTTR) family that regulates the transcription of genes encoding the tricarboxylic acid branch enzymes of the TCA cycle by responding to a pathway-specific metabolite, citrate. The biological function of CcpC has been characterized several times, but the structural basis for the molecular function of CcpC remains elusive. Here, we report characterization of a full-length CcpC from Bacillus amyloliquefaciens (BaCcpC-FL) and a crystal structure of the C-terminal inducer-binding domain (IBD) complexed with citrate. BaCcpC required both dyad symmetric regions I and II to recognize the citB promoter, and the presence of citrate reduced citB promoter binding. The crystal structure of CcpC-IBD shows two subdomains, IBD-I and IBD-II, and a citrate molecule buried between them. Ile100, two arginines (Arg147 and Arg260), and three serines (Ser129, Ser189, and Ser191) have strong hydrogen-bond interactions with citrate molecules. A structural comparison of BaCcpC-IBD with its homologues shows that they share the same tail-to-tail dimer alignment, but the dimeric interface and the rotation between these molecules exhibit significant differences. In addition, citrate can convert large BaCcpC-FL oligomers to monomers in solution. Taken together, our results provide a framework for understanding the mechanism underlying the functional divergence of the CcpC protein.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
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