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Title: GlpR Is a Direct Transcriptional Repressor of Fructose Metabolic Genes in Haloferax volcanii

Abstract

DeoR-type helix-turn-helix (HTH) domain proteins are transcriptional regulators of sugar and nucleoside metabolism in diverse bacteria and also occur in select archaea. In the model archaeon Haloferax volcanii, previous work implicated GlpR, a DeoR-type transcriptional regulator, in the transcriptional repression of glpR and the gene encoding the fructose-specific phosphofructokinase ( pfkB) during growth on glycerol. However, the global regulon governed by GlpR remained unclear. Here, we compared transcriptomes of wild-type and ΔglpR mutant strains grown on glycerol and glucose to detect significant transcript level differences for nearly 50 new genes regulated by GlpR. By coupling computational prediction of GlpR binding sequences with in vivo and in vitro DNA binding experiments, we determined that GlpR directly controls genes encoding enzymes involved in fructose degradation, including fructose bisphosphate aldolase, a central control point in glycolysis. GlpR also directly controls other transcription factors. In contrast, other metabolic pathways appear to be under the indirect influence of GlpR. In vitro experiments demonstrated that GlpR purifies to function as a tetramer that binds the effector molecule fructose-1-phosphate (F1P). Furthermore, these results suggest that H. volcanii GlpR functions as a direct negative regulator of fructose degradation during growth on carbon sources other than fructose, such asmore » glucose and glycerol, and that GlpR bears striking functional similarity to bacterial DeoR-type regulators.« less

Authors:
 [1];  [1];  [2];  [1];  [3];  [1];  [1]; ORCiD logo [3]; ORCiD logo [1];  [4]
  1. Univ. of Florida, Gainesville, FL (United States)
  2. Univ. of Florida, Gainesville, FL (United States); Washington State Univ., Richland, WA (United States)
  3. Duke Univ., Durham, NC (United States)
  4. Univ. of Illinois at Urbana Champaign, Urbana, IL (United States)
Publication Date:
Research Org.:
Univ. of Florida, Gainesville, FL (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1541706
Grant/Contract Number:  
FG02-05ER15650
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Bacteriology
Additional Journal Information:
Journal Volume: 200; Journal Issue: 17; Journal ID: ISSN 0021-9193
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; microbiology; transcription factor; glycerol; fructose metabolism; catabolite repression; glucose metabolism; Archaea

Citation Formats

Martin, Jonathan H., Rawls, Katherine Sherwood, Chan, Jou Chin, Hwang, Sungmin, Martinez-Pastor, Mar, McMillan, Lana J., Prunetti, Laurence, Schmid, Amy K., Maupin-Furlow, Julie A., and Metcalf, William W. GlpR Is a Direct Transcriptional Repressor of Fructose Metabolic Genes in Haloferax volcanii. United States: N. p., 2018. Web. doi:10.1128/jb.00244-18.
Martin, Jonathan H., Rawls, Katherine Sherwood, Chan, Jou Chin, Hwang, Sungmin, Martinez-Pastor, Mar, McMillan, Lana J., Prunetti, Laurence, Schmid, Amy K., Maupin-Furlow, Julie A., & Metcalf, William W. GlpR Is a Direct Transcriptional Repressor of Fructose Metabolic Genes in Haloferax volcanii. United States. doi:10.1128/jb.00244-18.
Martin, Jonathan H., Rawls, Katherine Sherwood, Chan, Jou Chin, Hwang, Sungmin, Martinez-Pastor, Mar, McMillan, Lana J., Prunetti, Laurence, Schmid, Amy K., Maupin-Furlow, Julie A., and Metcalf, William W. Mon . "GlpR Is a Direct Transcriptional Repressor of Fructose Metabolic Genes in Haloferax volcanii". United States. doi:10.1128/jb.00244-18. https://www.osti.gov/servlets/purl/1541706.
@article{osti_1541706,
title = {GlpR Is a Direct Transcriptional Repressor of Fructose Metabolic Genes in Haloferax volcanii},
author = {Martin, Jonathan H. and Rawls, Katherine Sherwood and Chan, Jou Chin and Hwang, Sungmin and Martinez-Pastor, Mar and McMillan, Lana J. and Prunetti, Laurence and Schmid, Amy K. and Maupin-Furlow, Julie A. and Metcalf, William W.},
abstractNote = {DeoR-type helix-turn-helix (HTH) domain proteins are transcriptional regulators of sugar and nucleoside metabolism in diverse bacteria and also occur in select archaea. In the model archaeon Haloferax volcanii, previous work implicated GlpR, a DeoR-type transcriptional regulator, in the transcriptional repression of glpR and the gene encoding the fructose-specific phosphofructokinase (pfkB) during growth on glycerol. However, the global regulon governed by GlpR remained unclear. Here, we compared transcriptomes of wild-type and ΔglpR mutant strains grown on glycerol and glucose to detect significant transcript level differences for nearly 50 new genes regulated by GlpR. By coupling computational prediction of GlpR binding sequences with in vivo and in vitro DNA binding experiments, we determined that GlpR directly controls genes encoding enzymes involved in fructose degradation, including fructose bisphosphate aldolase, a central control point in glycolysis. GlpR also directly controls other transcription factors. In contrast, other metabolic pathways appear to be under the indirect influence of GlpR. In vitro experiments demonstrated that GlpR purifies to function as a tetramer that binds the effector molecule fructose-1-phosphate (F1P). Furthermore, these results suggest that H. volcanii GlpR functions as a direct negative regulator of fructose degradation during growth on carbon sources other than fructose, such as glucose and glycerol, and that GlpR bears striking functional similarity to bacterial DeoR-type regulators.},
doi = {10.1128/jb.00244-18},
journal = {Journal of Bacteriology},
number = 17,
volume = 200,
place = {United States},
year = {2018},
month = {6}
}

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