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rocG [2019-10-30 09:16:30]

trigger enzyme, glutamate dehydrogenase and effector protein for [protein|87BCAE725B02860156D50E1783F6DB68510C811E|GltC]

locus

BSU_37790

6.28

46.48 kDa

protein length

424 aa Sequence Blast

gene length

1275 bp Sequence Blast

function

arginine utilization, controls the activity of [protein|87BCAE725B02860156D50E1783F6DB68510C811E|GltC]

product

glutamate dehydrogenase

essential

1.4.1.2

synonyms

Outlinks

KEGG BsubCyc SubtiList UniProt Expression Browser

Genomic Context

Hide small RNAs Zoom:

categories

[category|SW 2|Metabolism] → [category|SW 2.3|Amino acid/ nitrogen metabolism] → [category|SW 2.3.2|Utilization of amino acids] → [category|SW 2.3.2.1|Utilization of glutamine/ glutamate]

[category|SW 2|Metabolism] → [category|SW 2.3|Amino acid/ nitrogen metabolism] → [category|SW 2.3.2|Utilization of amino acids] → [category|SW 2.3.2.4|Utilization of arginine/ ornithine]

[category|SW 3|Information processing] → [category|SW 3.4|Regulation of gene expression] → [category|SW 3.4.2|Transcription factors and their control] → [category|SW 3.4.2.7|Control of transcription factor (other than two-component system)]

[category|SW 3|Information processing] → [category|SW 3.4|Regulation of gene expression] → [category|SW 3.4.3|Trigger enzyme] → [category|SW 3.4.3.2|Trigger enzymes that control gene expression by protein-protein interaction with transcription factors]

Gene

Coordinates

3,880,740 3,882,014

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 ([gene|C36C8C9EEDCAD392D9BEC9728A1E0CBFF2F4E790|''gudB1'']]])

sensitive to -lactam antibiotics such as cefuroxime and to fosfomycin (suppressed by activation of ''[gene|C36C8C9EEDCAD392D9BEC9728A1E0CBFF2F4E790|gudB]'') due to the downregulation of the [SW|SigW regulon] [Pubmed|22178969]

transcription profile of a ''[gene|56CBEBCFEF5CFB4A0175498338C7AF2F45EAA3E3|rocG] [gene|C36C8C9EEDCAD392D9BEC9728A1E0CBFF2F4E790|gudB]'' mutant strain: [http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE34383&submit.x=22&submit.y=9 GEO] [Pubmed|22178969]

The protein

Catalyzed reaction/ biological activity

H₂O + L-glutamate + NAD⁺ --> 2-oxoglutarate + H⁺ + NADH + NH₄⁺ (according to UniProt)

controls the activity of the [protein|87BCAE725B02860156D50E1783F6DB68510C811E|GltC] transcription activator [Pubmed|17608797]

Protein family

Glu/Leu/Phe/Val dehydrogenases family (with [protein|A52E50104B18D0A8518218C52D9CC36FDDB29AAF|Bcd] and [protein|C36C8C9EEDCAD392D9BEC9728A1E0CBFF2F4E790|GudB], according to UniProt)

Paralogous protein(s)

[protein|C36C8C9EEDCAD392D9BEC9728A1E0CBFF2F4E790|GudB]

Kinetic information

K_M [glutamate] = 2.9 mM, K_M [ammonium] = 18 mM [Pubmed|20630473]

[SW|Cofactors]

NAD⁺/NADH + H⁺

Structure

[PDB|3K92] (super-repressor mutant that is capable of constitutive inactivation of [protein|87BCAE725B02860156D50E1783F6DB68510C811E|GltC], E93K mutation) [Pubmed|20630473]

Expression and Regulation

Operons

genes

[gene|56CBEBCFEF5CFB4A0175498338C7AF2F45EAA3E3|rocG]

description

[Pubmed|10468601]

sigma factors

regulatory mechanism

regulation

expression during spore [SW|germination] is strongly reduced under conditions of osmotic stress [Pubmed|27766092]

additional information

view in new tab

Biological materials

Mutant

GP747 (spc), GP726 (aphA3), GP810 (del tet), GP1157 (cat) all available in [SW|Jörg Stülke]'s lab

BKE37790 ([gene|56CBEBCFEF5CFB4A0175498338C7AF2F45EAA3E3|rocG]::erm trpC2) available at [http://www.bgsc.org/getdetail.php?bgscid=BKE37790 BGSC], [Pubmed|28189581], upstream reverse: _UP1_CATCTTTTTCACCTCATTGT, downstream forward: _UP4_TAATTTGAGAAGCCTCCGCA

BKK37790 ([gene|56CBEBCFEF5CFB4A0175498338C7AF2F45EAA3E3|rocG]::kan trpC2) available at [http://www.bgsc.org/getdetail.php?bgscid=BKK37790 BGSC], [Pubmed|28189581], upstream reverse: _UP1_CATCTTTTTCACCTCATTGT, downstream forward: _UP4_TAATTTGAGAAGCCTCCGCA

Expression vectors

expression of native ''rocG'' in ''B. subtilis'': pGP529 (in [SW|pBQ200]), available in [SW|Jörg Stülke]'s lab [Pubmed|18326565]

for purification of RocG from ''E. coli'' carrying an N-terminal Strep-tag: pGP902 (in [SW|pGP172]), a series of ''rocG'' variants is also available in [SW|pGP172], available in [SW|Jörg Stülke]'s lab

for expression/ purification from ''E. coli'' with N-terminal His-tag and thrombin cleavage site, in [SW|pWH844]: pGP860, available in [SW|Jörg Stülke]'s lab

purification from ''B. subtilis'' with an N-terminal Strep-tag, for [SW|SPINE], (in [SW|pGP380]): pGP1709, available in [SW|Jörg Stülke]'s lab

two-hybrid system

''B. pertussis'' adenylate cyclase-based bacterial two hybrid system ([SW|BACTH]), available in [SW|Jörg Stülke]'s lab

Antibody

available in [SW|Jörg Stülke]'s lab [Pubmed|17183217]

labs

[SW|Linc Sonenshein], Tufts University, Boston, MA, USA [http://www.tufts.edu/sackler/microbiology/faculty/sonenshein/index.html Homepage]

[SW|Jörg Stülke], University of Göttingen, Germany [http://wwwuser.gwdg.de/~genmibio/stuelke.html Homepage]

[SW|Fabian Commichau] University of Göttingen, Germany [http://genmibio.uni-goettingen.de/index.php?id=130 Homepage]

References

Reviews

19698086,8299344,7705101,19895831,22625175,28615289

Enzymatic activity of RocG

18603778,16244435,16195607,18326565,9829940,20630473,21965396,25711804,28468957

Function in the control of [protein|87BCAE725B02860156D50E1783F6DB68510C811E|GltC] activity

15150225,17994626,17608797,17183217,20630473

Expression of [gene|56CBEBCFEF5CFB4A0175498338C7AF2F45EAA3E3|rocG]

12634342,15150224,10468601,9829940,22900538,20817675,25777015,25880922,27766092

Structural analysis of glutamate dehydrogenase

8263917,20630473

Bypass of [gene|search|rocG ]mutations

21219666,22178973,23785476

Additional publications

23338837,23419162,22178969,24473333,29242163