RocG

• 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
Gene expression levels in SubtiExpress: rocG
Interactions involving this protein in SubtInteract: RocG
Metabolic function and regulation of this protein in SubtiPathways: rocG
MW, pI	46.2 kDa, 6.28
Gene length, protein length	1272 bp, 424 amino acids
Immediate neighbours	rocA, sivA
Sequences	Protein DNA DNA_with_flanks
Genetic context This image was kindly provided by SubtiList
Expression at a glance   PubMed

Categories containing this gene/protein

utilization of amino acids, glutamate metabolism, 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]

• GEO entry: [3] (rocG gudB mutant vs. wild type)

The protein

Basic information/ Evolution

• Catalyzed reaction/ biological activity: L-glutamate + H₂O + NAD⁺ = 2-oxoglutarate + NH₃ + NADH + H⁺ (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: K_M [glutamate] = 2.9 mM, K_M [ammonium] = 18 mM PubMed

• Domains:

• Modification:

• Cofactor(s): NAD⁺/NADH + H⁺

• Effectors of protein activity:

• Interactions:
  • RocG-GltC, this interaction prevents transcription activation of the gltA-gltB operon by GltC PubMed

• Localization:

Database entries

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

• UniProt: P39633

• KEGG entry: [4]

• E.C. number: 1.4.1.2

Expression and regulation

• Operon: rocG PubMed

• Expression browser: rocG PubMed

• Sigma factor: SigL PubMed

• Regulation:
  • induced by arginine (RocR, AhrC), ornithine or proline PubMed
  • subject to carbon catabolite repression (CcpA) PubMed

• Regulatory mechanism:
  • RocR: transcription activation PubMed
  • AhrC: transcription activation PubMed
  • CcpA: transcription repression PubMed
  • AbrB: transcription repression PubMed

• 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 Jörg Stülke's lab

• Expression vector:
  • expression of native rocG in B. subtilis: pGP529 (in pBQ200), available in Jörg Stülke's lab PubMed
  • 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 Jörg Stülke's lab
  • for expression/ purification from E. coli with N-terminal His-tag and thrombin cleavage site, in pWH844: pGP860, available in Jörg Stülke's lab
  • purification from B. subtilis with an N-terminal Strep-tag, for SPINE, (in pGP380): pGP1709, available in Jörg Stülke's lab

• lacZ fusion:

• GFP fusion:

• two-hybrid system: B. pertussis adenylate cyclase-based bacterial two hybrid system (BACTH), available in Jörg Stülke's lab

• Antibody: available in Jörg Stülke's lab

Labs working on this gene/protein

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

Jörg Stülke, University of Göttingen, Germany Homepage

Fabian Commichau University of Göttingen, Germany Homepage

References

Expression of rocG

Bogumiła C Marciniak, Monika Pabijaniak, Anne de Jong, Robert Dűhring, Gerald Seidel, Wolfgang Hillen, Oscar P Kuipers
High- and low-affinity cre boxes for CcpA binding in Bacillus subtilis revealed by genome-wide analysis.
BMC Genomics: 2012, 13;401
[PubMed:22900538] [WorldCat.org] [DOI] (I e)

Onuma Chumsakul, Hiroki Takahashi, Taku Oshima, Takahiro Hishimoto, Shigehiko Kanaya, Naotake Ogasawara, Shu Ishikawa
Genome-wide binding profiles of the Bacillus subtilis transition state regulator AbrB and its homolog Abh reveals their interactive role in transcriptional regulation.
Nucleic Acids Res: 2011, 39(2);414-28
[PubMed:20817675] [WorldCat.org] [DOI] (I p)

Boris R Belitsky, Hyun-Jin Kim, Abraham L Sonenshein
CcpA-dependent regulation of Bacillus subtilis glutamate dehydrogenase gene expression.
J Bacteriol: 2004, 186(11);3392-8
[PubMed:15150224] [WorldCat.org] [DOI] (P p)

Naima Ould Ali, Josette Jeusset, Eric Larquet, Eric Le Cam, Boris Belitsky, Abraham L Sonenshein, Tarek Msadek, Michel Débarbouillé
Specificity of the interaction of RocR with the rocG-rocA intergenic region in Bacillus subtilis.
Microbiology (Reading): 2003, 149(Pt 3);739-750
[PubMed:12634342] [WorldCat.org] [DOI] (P p)

B R Belitsky, A L Sonenshein
An enhancer element located downstream of the major glutamate dehydrogenase gene of Bacillus subtilis.
Proc Natl Acad Sci U S A: 1999, 96(18);10290-5
[PubMed:10468601] [WorldCat.org] [DOI] (P p)

B R Belitsky, A L Sonenshein
Role and regulation of Bacillus subtilis glutamate dehydrogenase genes.
J Bacteriol: 1998, 180(23);6298-305
[PubMed:9829940] [WorldCat.org] [DOI] (P p)