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: sacY
MW, pI	32 kDa, 5.883
Gene length, protein length	840 bp, 280 aa
Immediate neighbours	sacX, gspA
Sequences	Protein DNA DNA_with_flanks
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

• BsubCyc: BSU38420

• 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

• BsubCyc: BSU38420

• 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)