Crh

• Description: "Catabolite repression HPr-like protein", Cofactor of the CcpA transcription factor
  
  

Gene name	crh
Synonyms	yvcM
Essential	no
Product	catabolite repression HPr-like protein
Function	catabolite repression
Metabolic function and regulation of this protein in SubtiPathways: Central C-metabolism
MW, pI	9,2 kDa, 4.70
Gene length, protein length	255 bp, 85 amino acids
Immediate neighbours	yvcL, yvcN
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: BSU34740

Database entries

• DBTBS entry: no entry

• SubtiList entry: [1]

The protein

Basic information/ Evolution

• Catalyzed reaction/ biological activity:

• Protein family: HPr family (according to Swiss-Prot) PtsH,HPr family

• Paralogous protein(s): HPr

Extended information on the protein

• Kinetic information:

• Domains: HPr domain (1–85)

• Modification: phosphorylation on Ser46 by HprK PubMed

• Cofactor(s):

• Effectors of protein activity:

• Interactions: Crh-GapA PubMed, Crh-CcpA PubMed, HprK-Crh PubMed

• Localization:

Database entries

• Structure: 2AK7 (dimeric Crh-Ser46-P), 1ZVV (CcpA-Crh-DNA complex), 2RLZ (dimer), 1MU4, NCBI, dimer NCBI, CcpA-Crh-DNA complex NCBI, dimeric phosphor-Crh NCBI
• Swiss prot entry: O06976

• KEGG entry: [2]

Additional information

Crh does not possess the phosphorylation site used for PTS phosphotransfer (His-15 in PtsH), it can only be phosphorylated on Ser-46

Expression and regulation

• Operon: yvcI-yvcJ-yvcK-yvcL-crh-yvcN PubMed

• Sigma factor: SigA PubMed

• Regulation: very weak stimuation of expression by citrate and succinate PubMed

• Regulatory mechanism:

• Additional information: Crh is weakly expressed. This results in part from a poorly conserved ribosomal binding site of the mRNA. PubMed

Biological materials

• Mutant: GP860 (aphA3), QB7097 (spc), available in Stülke lab

• Expression vector: pGP41 (N-terminal Strep-tag, purification from B. subtilis, for SPINE, in pGP380), pGP734 (C-terminal Strep-tag, purification from B. subtilis, for SPINE, in pGP382), available in Stülke lab

• lacZ fusion: see yvcI

• GFP fusion:

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

• Antibody: available in Stülke lab (not very good)

Labs working on this gene/protein

Boris Görke, University of Göttingen, Germany Homepage

Anne Galinier, University of Marseille, France

Wolfgang Hillen, Erlangen University, Germany Homepage

Richard Brennan, Houston, Texas, USA Homepage

References

1. Juy M, Penin F, Favier A (2003) Dimerization of Crh by reversible 3D domain swapping induces structural adjustments to its monomeric homologue Hpr. J Mol Biol. 332(4):767-76. PubMed
2. Galinier A, Deutscher J, Martin-Verstraete I: Phosphorylation of either Crh or HPr mediates binding of CcpA to the Bacillus subtilis xyn cre and catabolite repression of the xyn operon. J Mol Biol 1999, 286:307-314. PubMed
3. Galinier, A., Haiech, J., Kilhoffer, M.-C., Jaquinod, M., Stülke, J., Deutscher, J., & Martin-Verstraete, I. (1997) The Bacillus subtilis crh gene encodes a HPr-like protein involved in carbon catabolite repression. Proc. Natl. Acad. Sci. USA 94: 8439-8444. PubMed
4. Görke et al. (2005) YvcK of Bacillus subtilis is required for a normal cell shape and for growth on Krebs cycle intermediates and substrates of the pentose phosphate pathway. Microbiology 151: 3777-3791. PubMed
5. Görke, B., Fraysse, L. & Galinier, A. Drastic differences in Crh and HPr synthesis levels reflect their different impacts on catabolite repression in Bacillus subtilis. J. Bacteriol. 186, 2992-2995 (2004). PubMed
6. Martin-Verstraete, I., Deutscher, J., and Galinier, A. (1999) Phosphorylation of HPr and Crh by HprK, early steps in the catabolite repression signalling pathway for the Bacillus subtilis levanase operon. J Bacteriol 181: 2966-2969. PubMed
7. Pompeo et al. (2007) Interaction of GapA with HPr and its homologue, Crh: Novel levels of regulation of a key step of glycolysis in Bacillus subtilis? J Bacteriol 189, 1154-1157.PubMed
8. Schumacher, M. A., Seidel, G., Hillen, W. & Brennan, R. G. Phosphoprotein Crh-Ser46-P displays altered binding to CcpA to effect carbon catabolite regulation. J. Biol. Chem. 281, 6793-6800 (2006). PubMed