RocG

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  • Description: trigger enzyme: catabolic glutamate dehydrogenase induced by arginine, ornithine or proline, subject to carbon catabolite repression

Gene name rocG
Synonyms
Essential no
Product trigger enzyme: glutamate dehydrogenase (major)
Function arginine utilization, controls the activity of GltC
Interactions involving this protein in SubtInteract: RocG
Metabolic function and regulation of this protein in SubtiPathways:
Ammonium/ glutamate
MW, pI 46.2 kDa, 6.28
Gene length, protein length 1272 bp, 424 amino acids
Immediate neighbours rocA, yweA
Get the DNA and protein sequences
(Barbe et al., 2009)
Genetic context
RocG context.gif
This image was kindly provided by SubtiList








Categories containing this gene/protein

utilization of amino acids, transcription factors and their control, trigger enzyme

This gene is a member of the following regulons

AbrB regulon, AhrC regulon, CcpA regulon, RocR regulon, SigL regulon

The gene

Basic information

  • Locus tag: BSU37790

Phenotypes of a mutant

  • Poor growth on complex media such as SP (sporulation medium). No growth in minimal media with arginine as the only carbon source. Rapid accumulation of suppressor mutants (gudB1)
  • sensitive to ß-lactam antibiotics such as cefuroxime and to fosfomycin (suppressed by activation of gudB) due to the downregulation of the SigW regulon PubMed
  • transcription profile of a rocG gudB mutant strain: GEO PubMed

Database entries

  • DBTBS entry: [1]
  • SubtiList entry: [2]

Additional information

The protein

Basic information/ Evolution

  • Catalyzed reaction/ biological activity: L-glutamate + H2O + NAD+ = 2-oxoglutarate + NH3 + NADH (according to Swiss-Prot), controls the activity of the GltC transcription activator PubMed
  • Protein family: Glu/Leu/Phe/Val dehydrogenases family (according to Swiss-Prot)
  • Paralogous protein(s): GudB

Extended information on the protein

  • Kinetic information: KM [glutamate] = 2.9 mM, KM [ammonium] = 18 mM PubMed
  • Domains:
  • Modification:
  • Cofactor(s): NAD+/NADH2
  • Effectors of protein activity:

Database entries

  • Structure: 3K92 (super-repressor mutant that is capable of constitutive inactivation of GltC, E93K mutation) PubMed
  • KEGG entry: [4]

Additional information

Expression and regulation

  • Regulation:
  • Additional information:

Activation by RocR requires binding of RocR to a downstream element PubMed

Biological materials

  • Mutant: GP747 (spc), GP726 (aphA3), GP810 (del tet), GP1157 (cat) all available in Stülke lab
  • Expression vector:
    • expression of native rocG in B. subtilis: pGP529 (in pBQ200), available in Stülke lab
    • for purification of RocG from E. coli carrying an N-terminal Strep-tag: pGP902 (in pGP172), a series of rocG variants is also available in pGP172, available in Stülke lab
    • for expression/ purification from E. coli with N-terminal His-tag and thrombin cleavage site, in pWH844: pGP860, available in Stülke lab
    • purification from B. subtilis with an N-terminal Strep-tag, for SPINE, (in pGP380): pGP1709, available in Stülke lab
  • lacZ fusion:
  • GFP fusion:
  • two-hybrid system: B. pertussis adenylate cyclase-based bacterial two hybrid system (BACTH), available in Stülke lab
  • Antibody: available in Stülke lab

Labs working on this gene/protein

Linc Sonenshein, Tufts University, Boston, MA, USA Homepage

Jörg Stülke, University of Göttingen, Germany Homepage

Your additional remarks

References

Reviews

Enzymatic activity of RocG

Function in the control of GltC activity

Katrin Gunka, Joseph A Newman, Fabian M Commichau, Christina Herzberg, Cecilia Rodrigues, Lorraine Hewitt, Richard J Lewis, Jörg Stülke
Functional dissection of a trigger enzyme: mutations of the bacillus subtilis glutamate dehydrogenase RocG that affect differentially its catalytic activity and regulatory properties.
J Mol Biol: 2010, 400(4);815-27
[PubMed:20630473] [WorldCat.org] [DOI] (I p)

Christina Herzberg, Lope Andrés Flórez Weidinger, Bastian Dörrbecker, Sebastian Hübner, Jörg Stülke, Fabian M Commichau
SPINE: a method for the rapid detection and analysis of protein-protein interactions in vivo.
Proteomics: 2007, 7(22);4032-5
[PubMed:17994626] [WorldCat.org] [DOI] (P p)

Fabian M Commichau, Christina Herzberg, Philipp Tripal, Oliver Valerius, Jörg Stülke
A regulatory protein-protein interaction governs glutamate biosynthesis in Bacillus subtilis: the glutamate dehydrogenase RocG moonlights in controlling the transcription factor GltC.
Mol Microbiol: 2007, 65(3);642-54
[PubMed:17608797] [WorldCat.org] [DOI] (P p)

Fabian M Commichau, Ingrid Wacker, Jan Schleider, Hans-Matti Blencke, Irene Reif, Philipp Tripal, Jörg Stülke
Characterization of Bacillus subtilis mutants with carbon source-independent glutamate biosynthesis.
J Mol Microbiol Biotechnol: 2007, 12(1-2);106-13
[PubMed:17183217] [WorldCat.org] [DOI] (P p)

Boris R Belitsky, Abraham L Sonenshein
Modulation of activity of Bacillus subtilis regulatory proteins GltC and TnrA by glutamate dehydrogenase.
J Bacteriol: 2004, 186(11);3399-407
[PubMed:15150225] [WorldCat.org] [DOI] (P p)

Expression of rocG

Additional publications: PubMed

Structural analysis of glutamate dehydrogenase

Bypass of rocG mutations

Katrin Gunka, Stefan Tholen, Jan Gerwig, Christina Herzberg, Jörg Stülke, Fabian M Commichau
A high-frequency mutation in Bacillus subtilis: requirements for the decryptification of the gudB glutamate dehydrogenase gene.
J Bacteriol: 2012, 194(5);1036-44
[PubMed:22178973] [WorldCat.org] [DOI] (I p)

Lope A Flórez, Katrin Gunka, Rafael Polanía, Stefan Tholen, Jörg Stülke
SPABBATS: A pathway-discovery method based on Boolean satisfiability that facilitates the characterization of suppressor mutants.
BMC Syst Biol: 2011, 5;5
[PubMed:21219666] [WorldCat.org] [DOI] (I e)

Additional publications

PubMed

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