Difference between revisions of "SacT"

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* '''GFP fusion:'''
 
* '''GFP fusion:'''
 +
 +
* '''FLAG-tag construct:''' GP1223 (spc, based on [[pGP1331]]), available in the [[Stülke]] lab
  
 
* '''two-hybrid system:'''  
 
* '''two-hybrid system:'''  

Revision as of 13:23, 28 September 2012

Gene name sacT
Synonyms ipa-47d
Essential no
Product transcriptional antiterminator
Function regulation of sucrose utilization
Gene expression levels in SubtiExpress: sacT
Interactions involving this protein in SubtInteract: SacT
Metabolic function and regulation of this protein in SubtiPathways:
Sugar catabolism, Stress
MW, pI 31 kDa, 5.587
Gene length, protein length 828 bp, 276 aa
Immediate neighbours ywcJ, ywcI
Get the DNA and protein sequences
(Barbe et al., 2009)
Genetic context
SacT context.gif
This image was kindly provided by SubtiList
Expression at a glance   PubMed
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Categories containing this gene/protein

utilization of specific carbon sources, transcription factors and their control, RNA binding regulators, phosphoproteins

This gene is a member of the following regulons

The SacT regulon: sacP-sacA-ywdA

The gene

Basic information

  • Locus tag: BSU38070

Phenotypes of a mutant

Database entries

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

Additional information

The protein

Basic information/ Evolution

  • Catalyzed reaction/ biological activity: binding to the mRNA of the sacP-sacA operon, causes transcription antitermination (in presence of sucrose and absence of glucose)

Extended information on the protein

  • Kinetic information:
  • Domains:
  • Modification:
  • Cofactor(s):
  • Effectors of protein activity:

Database entries

  • Structure:
  • KEGG entry: [3]
  • E.C. number:

Additional information

Expression and regulation

  • Sigma factor:
  • Regulation:
    • repressed by casamino acids PubMed
  • Regulatory mechanism:
  • Additional information:

Biological materials

  • Mutant: GP429 (spc), available in Stülke lab
  • Expression vector:
    • for expression, purification of both PRDs in E. coli with N-terminal His-tag, in pWH844: pGP166, available in Stülke lab
    • for expression, purification of PRD-1 in E. coli with N-terminal His-tag, in pWH844: pGP426, available in Stülke lab
    • for expression, purification of PRD-2 in E. coli with N-terminal His-tag, in pWH844: pGP427, available in Stülke lab
    • for expression, purification of PRD-1 in E. coli with N-terminal His-tag and thrombin cleavage site, in pGP570: pGP439, available in Stülke lab
    • for expression, purification of PRD-2 in E. coli with N-terminal His-tag and thrombin cleavage site, in pGP570: pGP440, available in Stülke lab
    • for expression, purification of the RNA-binding domain in E. coli with N-terminal His-tag and thrombin cleavage site, in pGP570: pGP571, available in Stülke lab
    • for expression of the RNA-binding domain in B. subtilis, in pBQ200: pGP446, available in Stülke lab
    • for expression, purification of sacT-full length in B. subtilis with C-terminal Strep-tag, in pGP382: pGP1064, available in Stülke lab
    • for expression, purification of sacT-full length in B. subtilis with N-terminal Strep-tag, in pGP380: pGP1068, available in Stülke lab
  • lacZ fusion:
  • GFP fusion:
  • FLAG-tag construct: GP1223 (spc, based on pGP1331), available in the Stülke lab
  • two-hybrid system:
  • Antibody:

Labs working on this gene/protein

Your additional remarks

References

Sebastian Hübner, Nathalie Declerck, Christine Diethmaier, Dominique Le Coq, Stephane Aymerich, Jörg Stülke
Prevention of cross-talk in conserved regulatory systems: identification of specificity determinants in RNA-binding anti-termination proteins of the BglG family.
Nucleic Acids Res: 2011, 39(10);4360-72
[PubMed:21278164] [WorldCat.org] [DOI] (I p)

Ulrike Mäder, Georg Homuth, Christian Scharf, Knut Büttner, Rüdiger Bode, Michael Hecker
Transcriptome and proteome analysis of Bacillus subtilis gene expression modulated by amino acid availability.
J Bacteriol: 2002, 184(15);4288-95
[PubMed:12107147] [WorldCat.org] [DOI] (P p)

M Arnaud, M Débarbouillé, G Rapoport, M H Saier, J Reizer
In vitro reconstitution of transcriptional antitermination by the SacT and SacY proteins of Bacillus subtilis.
J Biol Chem: 1996, 271(31);18966-72
[PubMed:8702561] [WorldCat.org] [DOI] (P p)

S Aymerich, M Steinmetz
Specificity determinants and structural features in the RNA target of the bacterial antiterminator proteins of the BglG/SacY family.
Proc Natl Acad Sci U S A: 1992, 89(21);10410-4
[PubMed:1279678] [WorldCat.org] [DOI] (P p)

M Arnaud, P Vary, M Zagorec, A Klier, M Debarbouille, P Postma, G Rapoport
Regulation of the sacPA operon of Bacillus subtilis: identification of phosphotransferase system components involved in SacT activity.
J Bacteriol: 1992, 174(10);3161-70
[PubMed:1577686] [WorldCat.org] [DOI] (P p)

M Debarbouille, M Arnaud, A Fouet, A Klier, G Rapoport
The sacT gene regulating the sacPA operon in Bacillus subtilis shares strong homology with transcriptional antiterminators.
J Bacteriol: 1990, 172(7);3966-73
[PubMed:2163394] [WorldCat.org] [DOI] (P p)