SubtiBank SubtiBank

Categories containing this gene/protein


biosynthesis/ acquisition of amino acids, glutamate metabolism, transcription factors and their control, trigger enzyme, phosphoproteins, most abundant proteins

This gene is a member of the following regulons


GlnR regulon, TnrA regulon

Gene


  • Coordinates on the chromosome (coding sequence): 1,878,425 -> 1,879,759
  • Phenotypes of a mutant

  • auxotrophic for glutamine
  • The protein


    Catalyzed reaction/ biological activity

  • ATP L-glutamate NH3 = ADP phosphate L-glutamine (according to Swiss-Prot)
  • Protein family

  • glutamine synthetase family (according to Swiss-Prot)
  • Paralogous protein(s)

    Kinetic information

  • K(M) for: Glu: 27 mM, ATP: 2.4 mM, ammonium: 0.18 mM; v(max): 3.7 µmol/min/mg
  • Domains

  • glutamate binding flap (aa 300 ... 306: protects unstable intermediates from abberant hydrolysis)
  • Modification

  • phosphorylated on ser/ thr/ tyr PubMed
  • in vitro phosphorylated by PrkC on Thr-26, Thr-147, Ser-207, and Thr-286 PubMed
  • Cofactors

  • Mg(2 )
  • Effectors of protein activity

  • feedback inhibition by glutamine, glutamine binds the entrance site for glutamate
  • activity is inhibited upon interaction with TnrA PubMed
  • Structure

  • 4LNN (apo-GS) PubMed
  • 3QAJ (complex with ATP)
  • 4S0R (the TnrA-GlnA complex) PubMed
  • A general discussion of GS structure
  • Localization

  • cytoplasm (according to Swiss-Prot)
  • Interactions

  • TnrA-GlnA PubMed
  • GlnR-GlnA PubMed, (only the feedback-inhibited enzyme interacts with TnrA and GlnR)
  • GlnA-NrgB PubMed
  • forms dodecamers PubMed
  • Additional information

  • GlnA is a homooligomer of 12 subunits
  • Expression and Regulation


    Operon

  • glnR-glnA PubMed
  • Sigma factor

  • SigA PubMed
  • Regulation

  • expressed in the absence of glutamine (GlnR) PubMed
  • repressed in the absence of good nitrogen sources (glutamine or ammonium) (TnrA) PubMed
  • Regulatory mechanism

  • GlnR-GlnA complex: transcription repression PubMed
  • TnrA: transcription repression PubMed
  • Additional information

  • belongs to the 100 most abundant proteins PubMed
  • the mRNA is cleaved by RNase III PubMed
  • Biological materials


    Mutant

  • GP247 (glnA::cat), available in Jörg Stülke's lab
  • BKE17460 (glnA::ermC) (available at the BGSC and in Jörg Stülke's lab)
  • GP2263 (glnA::ermC) (available in Jörg Stülke's lab)
  • BP148 (del(glnR-glnA)::cat), available in Fabian Commichau's lab
  • GP1883 (del(glnR-glnA)::ermC), available in Fabian Commichaus and Jörg Stülkes labs
  • Expression vector

  • expression/ purification from E. coli, with N-terminal Strep-tag (in pGP172): pGP174, available in Jörg Stülke's lab
  • pGP177 (N-terminal Strep-tag, purification from B. subtilis, for SPINE, in pBQ200), available in Jörg Stülke's lab
  • lacZ fusion

  • glnR-lacZ: pGP189 (in pAC7), available in Jörg Stülke's lab
  • GFP fusion

    two-hybrid system

    FLAG-tag construct

    Antibody

  • available in Karl Forchhammer's lab
  • Labs working on this gene/protein


  • Susan Fisher, Boston, USA homepage
  • References


    Reviews

    Gunka K, Commichau FM

    Control of glutamate homeostasis in Bacillus subtilis: a complex interplay between ammonium assimilation, glutamate biosynthesis and degradation

    Mol Microbiol. 2012 Jul;85(2):213-24. doi: 10.1111/j.1365-2958.2012.08105.x. Review. PubMed PMID: 22625175.
    Commichau FM, Stülke J

    Trigger enzymes: bifunctional proteins active in metabolism and in controlling gene expression

    Mol Microbiol. 2008 Feb;67(4):692-702. Review. PubMed PMID: 18086213.
    Fisher SH

    Regulation of nitrogen metabolism in Bacillus subtilis: vive la différence

    Mol Microbiol. 1999 Apr;32(2):223-32. Review. PubMed PMID: 10231480.

    Original publications

    DiChiara JM, Liu B, Figaro S, Condon C, Bechhofer DH

    Mapping of internal monophosphate 5' ends of Bacillus subtilis messenger RNAs and ribosomal RNAs in wild-type and ribonuclease-mutant strains

    Nucleic Acids Res. 2016 Apr 20;44(7):3373-89. doi: 10.1093/nar/gkw073. PubMed PMID: 26883633; PubMed Central PMCID: PMC4838370.
    Hauf K, Kayumov A, Gloge F, Forchhammer K

    The Molecular Basis of TnrA Control by Glutamine Synthetase in Bacillus subtilis

    J Biol Chem. 2016 Feb 12;291(7):3483-95. doi: 10.1074/jbc.M115.680991. PubMed PMID: 26635369; PubMed Central PMCID: PMC4751389.
    Mirouze N, Bidnenko E, Noirot P, Auger S

    Genome-wide mapping of TnrA-binding sites provides new insights into the TnrA regulon in Bacillus subtilis

    Microbiologyopen. 2015 Jun;4(3):423-35. doi: 10.1002/mbo3.249. PubMed PMID: 25755103; PubMed Central PMCID: PMC4475385.
    Schumacher MA, Chinnam NB, Cuthbert B, Tonthat NK, Whitfill T

    Structures of regulatory machinery reveal novel molecular mechanisms controlling B

    subtilis nitrogen homeostasis. Genes Dev. 2015 Feb 15;29(4):451-64. doi: 10.1101/gad.254714.114. PubMed PMID: 25691471; PubMed Central PMCID: PMC4335299.
    Murray DS, Chinnam N, Tonthat NK, Whitfill T, Wray LV Jr, Fisher SH, Schumacher MA

    Structures of the Bacillus subtilis glutamine synthetase dodecamer reveal large intersubunit catalytic conformational changes linked to a unique feedback inhibition mechanism

    J Biol Chem. 2013 Dec 13;288(50):35801-11. doi: 10.1074/jbc.M113.519496. PubMed PMID: 24158439; PubMed Central PMCID: PMC3861631.
    Fedorova K, Kayumov A, Woyda K, Ilinskaja O, Forchhammer K

    Transcription factor TnrA inhibits the biosynthetic activity of glutamine synthetase in Bacillus subtilis

    FEBS Lett. 2013 May 2;587(9):1293-8. doi: 10.1016/j.febslet.2013.03.015. PubMed PMID: 23535029.
    Kayumov A, Heinrich A, Fedorova K, Ilinskaya O, Forchhammer K

    Interaction of the general transcription factor TnrA with the PII-like protein GlnK and glutamine synthetase in Bacillus subtilis

    FEBS J. 2011 May;278(10):1779-89. doi: 10.1111/j.1742-4658.2011.08102.x. PubMed PMID: 21435182.
    Wray LV Jr, Fisher SH

    Functional roles of the conserved Glu304 loop of Bacillus subtilis glutamine synthetase

    J Bacteriol. 2010 Oct;192(19):5018-25. doi: 10.1128/JB.00509-10. PubMed PMID: 20656908; PubMed Central PMCID: PMC2944536.
    Pietack N, Becher D, Schmidl SR, Saier MH, Hecker M, Commichau FM, Stülke J

    In vitro phosphorylation of key metabolic enzymes from Bacillus subtilis: PrkC phosphorylates enzymes from different branches of basic metabolism

    J Mol Microbiol Biotechnol. 2010;18(3):129-40. doi: 10.1159/000308512. PubMed PMID: 20389117.
    Fisher SH, Wray LV Jr

    Novel trans-Acting Bacillus subtilis glnA mutations that derepress glnRA expression

    J Bacteriol. 2009 Apr;191(8):2485-92. doi: 10.1128/JB.01734-08. PubMed PMID: 19233925; PubMed Central PMCID: PMC2668423.
    Fisher SH, Wray LV Jr

    Novel trans-Acting Bacillus subtilis glnA mutations that derepress glnRA expression

    J Bacteriol. 2009 Apr;191(8):2485-92. doi: 10.1128/JB.01734-08. PubMed PMID: 19233925; PubMed Central PMCID: PMC2668423.
    Wray LV Jr, Fisher SH

    Bacillus subtilis GlnR contains an autoinhibitory C-terminal domain required for the interaction with glutamine synthetase

    Mol Microbiol. 2008 Apr;68(2):277-85. doi: 10.1111/j.1365-2958.2008.06162.x. PubMed PMID: 18331450.
    Fisher SH, Wray LV Jr

    Bacillus subtilis glutamine synthetase regulates its own synthesis by acting as a chaperone to stabilize GlnR-DNA complexes

    Proc Natl Acad Sci U S A. 2008 Jan 22;105(3):1014-9. doi: 10.1073/pnas.0709949105. PubMed PMID: 18195355; PubMed Central PMCID: PMC2242682.
    Fisher SH, Wray LV Jr

    Feedback-resistant mutations in Bacillus subtilis glutamine synthetase are clustered in the active site

    J Bacteriol. 2006 Aug;188(16):5966-74. PubMed PMID: 16885465; PubMed Central PMCID: PMC1540052.
    Zalieckas JM, Wray LV Jr, Fisher SH

    Cross-regulation of the Bacillus subtilis glnRA and tnrA genes provides evidence for DNA binding site discrimination by GlnR and TnrA

    J Bacteriol. 2006 Apr;188(7):2578-85. PubMed PMID: 16547045; PubMed Central PMCID: PMC1428417.
    Lévine A, Vannier F, Absalon C, Kuhn L, Jackson P, Scrivener E, Labas V, Vinh J, Courtney P, Garin J, Séror SJ

    Analysis of the dynamic Bacillus subtilis Ser/Thr/Tyr phosphoproteome implicated in a wide variety of cellular processes

    Proteomics. 2006 Apr;6(7):2157-73. PubMed PMID: 16493705.
    Wray LV Jr, Fisher SH

    A feedback-resistant mutant of Bacillus subtilis glutamine synthetase with pleiotropic defects in nitrogen-regulated gene expression

    J Biol Chem. 2005 Sep 30;280(39):33298-304. PubMed PMID: 16055443.
    Eymann C, Dreisbach A, Albrecht D, Bernhardt J, Becher D, Gentner S, Tam le T, Büttner K, Buurman G, Scharf C, Venz S, Völker U, Hecker M

    A comprehensive proteome map of growing Bacillus subtilis cells

    Proteomics. 2004 Oct;4(10):2849-76. PubMed PMID: 15378759.
    Fisher SH, Brandenburg JL, Wray LV Jr

    Mutations in Bacillus subtilis glutamine synthetase that block its interaction with transcription factor TnrA

    Mol Microbiol. 2002 Aug;45(3):627-35. PubMed PMID: 12139611.
    Wray LV Jr, Ferson AE, Rohrer K, Fisher SH

    TnrA, a transcription factor required for global nitrogen regulation in Bacillus subtilis

    Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):8841-5. PubMed PMID: 8799114; PubMed Central PMCID: PMC38555.
    Brown SW, Sonenshein AL

    Autogenous regulation of the Bacillus subtilis glnRA operon

    J Bacteriol. 1996 Apr;178(8):2450-4. PubMed PMID: 8636055; PubMed Central PMCID: PMC177962.
    Schreier HJ, Rostkowski CA, Kellner EM

    Altered regulation of the glnRA operon in a Bacillus subtilis mutant that produces methionine sulfoximine-tolerant glutamine synthetase

    J Bacteriol. 1993 Feb;175(3):892-7. PubMed PMID: 8093698; PubMed Central PMCID: PMC196239.
    Fisher SH, Sonenshein AL

    Bacillus subtilis glutamine synthetase mutants pleiotropically altered in glucose catabolite repression

    J Bacteriol. 1984 Feb;157(2):612-21. PubMed PMID: 6141156; PubMed Central PMCID: PMC215290.
    Strauch MA, Aronson AI, Brown SW, Schreier HJ, Sonenhein AL

    Sequence of the Bacillus subtilis glutamine synthetase gene region

    Gene. 1988 Nov 30;71(2):257-65. PubMed PMID: 2906311.
    Schreier HJ, Brown SW, Hirschi KD, Nomellini JF, Sonenshein AL

    Regulation of Bacillus subtilis glutamine synthetase gene expression by the product of the glnR gene

    J Mol Biol. 1989 Nov 5;210(1):51-63. Erratum in: J Mol Biol 1992 Jul 20;226(2):571. PubMed PMID: 2573733.
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