SacY

• Description: transcriptional antiterminator for sacB and sacX-sacY (acts at high sucrose concentrations)
  
  

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 This image was kindly provided by SubtiList
Expression at a glance   PubMed

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 gene

Basic information

• Locus tag: BSU38420

Database entries

• DBTBS entry: [1]

• SubtiList entry: [2]

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)

• Protein family: transcriptional antiterminator bglG family (according to Swiss-Prot) BglG family of antiterminators

• Paralogous protein(s): LicT, GlcT, SacT

Extended information on the protein

• Kinetic information:
  • K(D) for the RAT-RNA: 3 nM PubMed
• Domains:

• Modification:

• Cofactor(s):

• Effectors of protein activity:

• Interactions:
  • PtsH-SacY

• Localization:

Database entries

• Structure: 1AUU (RNA-binding domain)

• UniProt: P15401

• KEGG entry: [3]

• E.C. number:

Expression and regulation

• Operon: sacX-sacY PubMed

• Expression browser: sacY PubMed

• Sigma factor: SigA PubMed

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

• Regulatory mechanism:
  • induction by SacY-dependent RNA switch (transcriptional antitermination) PubMed
  • DegU-P: transcription activation 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

References

Caroline Clerte, Nathalie Declerck, Emmanuel Margeat
Competitive folding of anti-terminator/terminator hairpins monitored by single molecule FRET.
Nucleic Acids Res: 2013, 41(4);2632-43
[PubMed:23303779] [WorldCat.org] [DOI] (I p)

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)

Nathalie Declerck, Nguyet Le Minh, Yinshang Yang, Vanessa Bloch, Michel Kochoyan, Stéphane Aymerich
RNA recognition by transcriptional antiterminators of the BglG/SacY family: mapping of SacY RNA binding site.
J Mol Biol: 2002, 319(5);1035-48
[PubMed:12079345] [WorldCat.org] [DOI] (P 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)