Difference between revisions of "SacY"

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Line 139: Line 139:
 
* '''lacZ fusion:'''
 
* '''lacZ fusion:'''
  
* '''GFP fusion:'''
+
* '''GFP fusion:''' GP1226 (spc, based on [[pGP1870]]), available in the [[Stülke]] lab
 +
 
 +
* '''YFP fusion:''' GP1230 (spc, based on [[pGP1871]]), available in the [[Stülke]] lab
  
 
* '''FLAG-tag construct:''' GP1222 (spc, based on [[pGP1331]]), available in the [[Stülke]] lab
 
* '''FLAG-tag construct:''' GP1222 (spc, based on [[pGP1331]]), available in the [[Stülke]] lab

Revision as of 13:34, 28 September 2012

Gene name sacY
Synonyms ipa-13r, sacS
Essential no
Product transcriptional antiterminator
Function regulation of sucrose utilization
Gene expression levels in SubtiExpress: sacY
Interactions involving this protein in SubtInteract: SacY
Metabolic function and regulation of this protein in SubtiPathways:
Sugar catabolism, Stress
MW, pI 32 kDa, 5.883
Gene length, protein length 840 bp, 280 aa
Immediate neighbours sacX, gspA
Get the DNA and protein sequences
(Barbe et al., 2009)
Genetic context
SacY context.gif
This image was kindly provided by SubtiList
Expression at a glance   PubMed
SacY expression.png




























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

SacY regulon

The SacY regulon: sacB-levB-yveA, sacX-sacY

The gene

Basic information

  • Locus tag: BSU38420

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 sacB and the sacX-sacY operon, causes transcription antitermination (in presence of sucrose)

Extended information on the protein

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

Database entries

  • Structure: 1AUU (RNA-binding domain)
  • KEGG entry: [3]
  • E.C. number:

Additional information

Expression and regulation

  • Regulation: induction by sucrose (at high concentration) PubMed
  • Additional information:

Biological materials

  • Mutant: GP425 (cat), available in Stülke lab
  • Expression vector:
    • for expression, purification of the RNA-binding domain in E. coli with N-terminal His-tag, in pWH844: pGP316, 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: pGP573, available in Stülke lab
  • lacZ fusion:
  • GFP fusion: GP1226 (spc, based on pGP1870), available in the Stülke lab
  • YFP fusion: GP1230 (spc, based on pGP1871), available in the Stülke lab
  • FLAG-tag construct: GP1222 (spc, based on pGP1331), available in the Stülke lab
  • two-hybrid system:
  • Antibody:

Labs working on this gene/protein

Stephane Aymerich, Microbiology and Molecular Genetics, INRA Paris-Grignon, France

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)

P Tortosa, N Declerck, H Dutartre, C Lindner, J Deutscher, D Le Coq
Sites of positive and negative regulation in the Bacillus subtilis antiterminators LicT and SacY.
Mol Microbiol: 2001, 41(6);1381-93
[PubMed:11580842] [WorldCat.org] [DOI] (P p)

N Declerck, F Vincent, F Hoh, S Aymerich, H van Tilbeurgh
RNA recognition by transcriptional antiterminators of the BglG/SacY family: functional and structural comparison of the CAT domain from SacY and LicT.
J Mol Biol: 1999, 294(2);389-402
[PubMed:10610766] [WorldCat.org] [DOI] (P p)

H van Tilbeurgh, X Manival, S Aymerich, J M Lhoste, C Dumas, M Kochoyan
Crystal structure of a new RNA-binding domain from the antiterminator protein SacY of Bacillus subtilis.
EMBO J: 1997, 16(16);5030-6
[PubMed:9305644] [WorldCat.org] [DOI] (P p)

X Manival, Y Yang, M P Strub, M Kochoyan, M Steinmetz, S Aymerich
From genetic to structural characterization of a new class of RNA-binding domain within the SacY/BglG family of antiterminator proteins.
EMBO J: 1997, 16(16);5019-29
[PubMed:9305643] [WorldCat.org] [DOI] (P p)

P Tortosa, S Aymerich, C Lindner, M H Saier, J Reizer, D Le Coq
Multiple phosphorylation of SacY, a Bacillus subtilis transcriptional antiterminator negatively controlled by the phosphotransferase system.
J Biol Chem: 1997, 272(27);17230-7
[PubMed:9202047] [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)

P Tortosa, D Le Coq
A ribonucleic antiterminator sequence (RAT) and a distant palindrome are both involved in sucrose induction of the Bacillus subtilis sacXY regulatory operon.
Microbiology (Reading): 1995, 141 ( Pt 11);2921-7
[PubMed:8535520] [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)

A M Crutz, M Steinmetz
Transcription of the Bacillus subtilis sacX and sacY genes, encoding regulators of sucrose metabolism, is both inducible by sucrose and controlled by the DegS-DegU signalling system.
J Bacteriol: 1992, 174(19);6087-95
[PubMed:1400159] [WorldCat.org] [DOI] (P p)

M M Zukowski, L Miller, P Cosgwell, K Chen, S Aymerich, M Steinmetz
Nucleotide sequence of the sacS locus of Bacillus subtilis reveals the presence of two regulatory genes.
Gene: 1990, 90(1);153-5
[PubMed:2116367] [WorldCat.org] [DOI] (P p)

A M Crutz, M Steinmetz, S Aymerich, R Richter, D Le Coq
Induction of levansucrase in Bacillus subtilis: an antitermination mechanism negatively controlled by the phosphotransferase system.
J Bacteriol: 1990, 172(2);1043-50
[PubMed:2105292] [WorldCat.org] [DOI] (P p)