The LacI family protein GlyR3 co-regulates the celC operon and manB in Clostridium thermocellum

Choi, Jinlyung; Klingeman, Dawn M.; Brown, Steven D.; Cox, Chris D.

doi:10.1186/s13068-017-0849-2

Title: The LacI family protein GlyR3 co-regulates the celC operon and manB in Clostridium thermocellum

Abstract

In this paper, we demonstrate that the GlyR3 protein mediates the regulation of manB. We first identify putative GlyR3 binding sites within or just upstream of the coding regions of manB and celT. Using an electrophoretic mobility shift assay (EMSA), we determined that a higher concentration of GlyR3 is required to effectively bind to the putative manB site in comparison to the celC site. Neither the putative celT site nor random DNA significantly binds GlyR3. While laminaribiose interfered with GlyR3 binding to the celC binding site, binding to the manB site was unaffected. In the presence of laminaribiose, in vivo transcription of the celC–glyR3–licA gene cluster increases, while manB expression is repressed, compared to in the absence of laminaribiose, consistent with the results from the EMSA. An in vitro transcription assay demonstrated that GlyR3 and laminaribiose interactions were responsible for the observed patters of in vivo transcription.

Authors:: Choi, Jinlyung; Klingeman, Dawn M.; Brown, Steven D.; Cox, Chris D.

Publication Date:: Sat Jun 24 00:00:00 EDT 2017

Research Org.:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Org.:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

OSTI Identifier:: 1618679

Alternate Identifier(s):: OSTI ID: 1376552

Grant/Contract Number:: AC05-00OR22725

Resource Type:: Published Article

Journal Name:: Biotechnology for Biofuels

Additional Journal Information:: Journal Name: Biotechnology for Biofuels Journal Volume: 10 Journal Issue: 1; Journal ID: ISSN 1754-6834

Publisher:: Springer Science + Business Media

Country of Publication:: Netherlands

Language:: English

Subject:: 09 BIOMASS FUELS; Clostridium thermocellum

Citation Formats


                    Choi, Jinlyung, Klingeman, Dawn M., Brown, Steven D., and Cox, Chris D. The LacI family protein GlyR3 co-regulates the celC operon and manB in Clostridium thermocellum.  Netherlands: N. p., 2017. 
Web.  doi:10.1186/s13068-017-0849-2.

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                    Choi, Jinlyung, Klingeman, Dawn M., Brown, Steven D., & Cox, Chris D. The LacI family protein GlyR3 co-regulates the celC operon and manB in Clostridium thermocellum.  Netherlands.  https://doi.org/10.1186/s13068-017-0849-2

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                    Choi, Jinlyung, Klingeman, Dawn M., Brown, Steven D., and Cox, Chris D. Sat .  
"The LacI family protein GlyR3 co-regulates the celC operon and manB in Clostridium thermocellum".  Netherlands.  https://doi.org/10.1186/s13068-017-0849-2.

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@article{osti_1618679,

  title        = {The LacI family protein GlyR3 co-regulates the celC operon and manB in Clostridium thermocellum},

  author       = {Choi, Jinlyung and Klingeman, Dawn M. and Brown, Steven D. and Cox, Chris D.},

  abstractNote = {In this paper, we demonstrate that the GlyR3 protein mediates the regulation of manB. We first identify putative GlyR3 binding sites within or just upstream of the coding regions of manB and celT. Using an electrophoretic mobility shift assay (EMSA), we determined that a higher concentration of GlyR3 is required to effectively bind to the putative manB site in comparison to the celC site. Neither the putative celT site nor random DNA significantly binds GlyR3. While laminaribiose interfered with GlyR3 binding to the celC binding site, binding to the manB site was unaffected. In the presence of laminaribiose, in vivo transcription of the celC–glyR3–licA gene cluster increases, while manB expression is repressed, compared to in the absence of laminaribiose, consistent with the results from the EMSA. An in vitro transcription assay demonstrated that GlyR3 and laminaribiose interactions were responsible for the observed patters of in vivo transcription.},

  doi          = {10.1186/s13068-017-0849-2},

  journal      = {Biotechnology for Biofuels},

  number       = 1,

  volume       = 10,

  place        = {Netherlands},

  year         = {Sat Jun 24 00:00:00 EDT 2017},

  month        = {Sat Jun 24 00:00:00 EDT 2017}

}

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