CitZ

• Description: citrate synthase
  
  

Gene name	citZ
Synonyms	citA2
Essential	no
Product	citrate synthase II
Function	TCA cycle
Metabolic function and regulation of this protein in SubtiPathways: Central C-metabolism
MW, pI	41 kDa, 5.451
Gene length, protein length	1116 bp, 372 aa
Immediate neighbours	icd, ytwI
Get the DNA and protein sequences (Barbe et al., 2009)
Genetic context This image was kindly provided by SubtiList

The gene

Basic information

• Locus tag: BSU29140

Phenotypes of a mutant

glutamate auxotrophy and a defect in sporulation PubMed

Database entries

• DBTBS entry: [1]

• SubtiList entry: [2]

The protein

Basic information/ Evolution

• Catalyzed reaction/ biological activity: Acetyl-CoA + H₂O + oxaloacetate = citrate + CoA (according to Swiss-Prot)

• Protein family: citrate synthase family (according to Swiss-Prot)

• Paralogous protein(s): MmgD, CitA

Extended information on the protein

• Kinetic information: Michaelis-Menten (Random Sequential Reaction Mechanism) PubMed

• Domains:

• Modification: phosphorylation on Ser-284 PubMed

• Cofactor(s):

• Effectors of protein activity:
  • Inhibited by acetyl-CoA, 2-oxoglutarate and NADH PubMed FEBS Letters
  • Inhibited by citrate and CoA (competitively against acetyl-CoA and non-competitively against oxaloacetate) PubMed
  • Inhibited by ATP competitively in B. subtilis strain 168 and HS 1A17 PubMed PubMed
    • In B. subtilis strain HS 2A2, ATP inhibits a non-competitive fashion PubMed
  • Activated by AMP PubMed

• Interactions:
  • CitZ-Mdh-Icd (this is the core of the TCA cycle metabolon) PubMed

• Localization:

Database entries

• Structure: 2C6X

• UniProt: P39120

• KEGG entry: [3]

• E.C. number: 2.3.3.1

Expression and regulation

• Operon:
  • citZ-icd-mdh PubMed

• Sigma factor: SigA PubMed

• Regulation:
  • repression by glucose (6.7-fold) (CcpA) PubMed
  • repression by glucose + glutamate (CcpC) PubMed

• Regulatory mechanism:
  • CcpA: transcription repression, CcpC: transcription repression PubMed
  • CcpC: transcription repression (molecular inducer: citrate) PubMed

• Additional information:
  • The mRNA has a long 5' leader region. This may indicate RNA-based regulation PubMed

Biological materials

• Mutant: GP678 (erm), available in Stülke lab

• Expression vector:
  • pGP1120 (N-terminal Strep-tag, for SPINE, purification from B. subtilis, in pGP380) (available in Stülke lab)
  • pGP1776 (for expression, purification in E. coli with N-terminal Strep-tag, in pGP172, available in Stülke lab)

• lacZ fusion:

• GFP fusion:

• two-hybrid system: B. pertussis adenylate cyclase-based bacterial two hybrid system (BACTH), available in Stülke lab

• Antibody: available in Linc Sonenshein lab

Labs working on this gene/protein

Linc Sonenshein, Tufts University, Boston, MA, USA Homepage

Jörg Stülke, University of Göttingen, Germany Homepage

References

Reviews

G Wiegand, S J Remington
Citrate synthase: structure, control, and mechanism.
Annu Rev Biophys Biophys Chem: 1986, 15;97-117
[PubMed:3013232] [WorldCat.org] [DOI] (P p)