Revision as of 04:24, 12 April 2009

Description: Enzyme I, general (non sugar-specific) component of the PTS. Enzyme I transfers the phosphoryl group from phosphoenolpyruvate (PEP) to the phosphoryl carrier protein (HPr)

Gene name	ptsI
Synonyms
Essential	no
Product	phosphotransferase system (PTS) enzyme I
Function	PTS-dependent sugar transport
MW, pI	62,9 kDa, 4.59
Gene length, protein length	1710 bp, 570 amino acids
Immediate neighbours	ptsH, splA
Gene sequence (+200bp)	Protein sequence
Caution: The sequence for this gene in SubtiList contains errors
Genetic context This image was kindly provided by SubtiList

The gene

Basic information

Coordinates: 1458959 - 1460668

Phenotypes of a mutant

Database entries

DBTBS entry: [1]

SubtiList entry: [2]

Additional information

The protein

Basic information/ Evolution

Catalyzed reaction/ biological activity: PEP-dependent autophosphorylation on His-189, transfer of the phosphoryl group to HPr (His-15)

Protein family: PEP-utilizing enzyme family PEP-utilizing enzyme family

Paralogous protein(s):

Extended information on the protein

Kinetic information:

Domains:
- HPr binding site (N-Terminal Domain)
- pyruvate binding site (C-Terminal Domain)
- pyrophosphate/phosphate carrier histidine (central Domain)

Modification: transient autophosphorylation on His-189, in vivo also phosphorylated on Ser-34 or Ser-36 PubMed

Cofactor(s): Magnesium

Effectors of protein activity:

Interactions:

Localization: Cytoplasm

Database entries

Structure:

Swiss prot entry: [3]

KEGG entry: [4]

E.C. number: 2.7.3.9

Additional information

Expression and regulation

Operon:
- ptsG-ptsH-ptsI
- ptsH-ptsI

Sigma factor: SigA PubMed
Regulation: expression activated by glucose (4.3 fold) PubMed, induction by glucose (ptsG), constitutive (ptsH)

Regulatory mechanism: ptsG: transcriptional antitermination via the GlcT-dependent RNA-switch

Additional information:

Biological materials

Mutant: GP864 (ermC), available in Stülke lab

Expression vector: pAG3 (His-tag), available in Galinier lab

lacZ fusion:

GFP fusion:

Antibody:

Labs working on this gene/protein

Josef Deutscher, Paris-Grignon, France

Jörg Stülke, University of Göttingen, Germany Homepage

Your additional remarks

References

Blencke et al. (2003) Transcriptional profiling of gene expression in response to glucose in Bacillus subtilis: regulation of the central metabolic pathways. Metab Eng. 5: 133-149 PubMed
Frisby, D., and Zuber, P. 1994. Mutations in pts cause catabolite-resistant sporulation and altered regulation of spo0H in Bacillus subtilis. J. Bacteriol. 176: 2587-2595. PubMed
Macek B, Mijakovic I, Olsen JV (2007) The serine/threonine/tyrosine phosphoproteome of the model bacterium Bacillus subtilis. Mol Cell Proteomics 6(4): 697-707. PubMed

@@ Line 1: / Line 1: @@
-* '''Description:''' HPr, General component of the sugar phosphotransferase system (PTS).  <br/><br/>
+* '''Description:''' Enzyme I, general (non sugar-specific) component of the PTS. Enzyme I transfers the phosphoryl group from phosphoenolpyruvate (PEP) to the phosphoryl carrier protein ([[PtsH |HPr]])  <br/><br/>
 {| align="right" border="1" cellpadding="2"
 |-
 |style="background:#ABCDEF;" align="center"|'''Gene name'''
-|''ptsH''
+|''ptsI''
 |-
 |style="background:#ABCDEF;" align="center"| '''Synonyms''' || '' ''
@@ Line 10: / Line 10: @@
 |style="background:#ABCDEF;" align="center"| '''Essential''' || no
 |-
-|style="background:#ABCDEF;" align="center"| '''Product''' || histidine-containing phosphocarrier <br/>protein HPr of the PTS
+|style="background:#ABCDEF;" align="center"| '''Product''' || phosphotransferase system (PTS) enzyme I
 |-
-|style="background:#ABCDEF;" align="center"|'''Function''' || PTS-dependent sugar transport <br/>and carbon catabolite repression
+|style="background:#ABCDEF;" align="center"|'''Function''' || PTS-dependent sugar transport
 |-
-|style="background:#ABCDEF;" align="center"| '''MW, pI''' || 9,1 kDa, 4.58
+|style="background:#ABCDEF;" align="center"| '''MW, pI''' || 62,9 kDa, 4.59
 |-
-|style="background:#ABCDEF;" align="center"| '''Gene length, protein length''' || 264 bp, 88 amino acids
+|style="background:#ABCDEF;" align="center"| '''Gene length, protein length''' || 1710 bp, 570 amino acids
 |-
-|style="background:#ABCDEF;" align="center"|'''Immediate neighbours''' || ''[[ptsG]]'', ''[[ptsI]]''
+|style="background:#ABCDEF;" align="center"|'''Immediate neighbours''' || ''[[ptsH]]'', ''[[splA]]''
 |-
-|style="background:#FAF8CC;" align="center"|'''[http://subtiwiki.uni-goettingen.de/ptsH_nucleotide.txt     Gene sequence      (+200bp)   ]'''
+|style="background:#FAF8CC;" align="center"|'''[http://subtiwiki.uni-goettingen.de/ptsI_nucleotide.txt     Gene sequence      (+200bp) ]'''
-|style="background:#FAF8CC;" align="center"|'''[http://subtiwiki.uni-goettingen.de/ptsH_protein.txt Protein sequence]'''
+|style="background:#FAF8CC;" align="center"|'''[http://subtiwiki.uni-goettingen.de/ptsI_protein.txt Protein sequence]'''
 |-
-|colspan="2" | '''Genetic context''' <br/> [[Image:ptsH_context.gif]]
+|colspan="2" style="background:#FAF8CC;color:#FF0000" align="center" | '''Caution: The sequence for this gene in SubtiList contains errors
+|-
+|colspan="2" | '''Genetic context''' <br/> [[Image:ptsI_context.gif]]
   <div align="right"> <small>This image was kindly provided by [http://genolist.pasteur.fr/SubtiList/ SubtiList]</small></div>
 |-
@@ Line 31: / Line 33: @@
 <br/><br/><br/><br/>
 =The gene=
@@ Line 37: / Line 38: @@
 === Basic information ===
-* '''Coordinates:''' 1458693 - 1458956
+* '''Coordinates:''' 1458959 - 1460668
 ===Phenotypes of a mutant ===
@@ Line 45: / Line 46: @@
 * '''DBTBS entry:''' [http://dbtbs.hgc.jp/COG/prom/ptsGHI.html]
-* '''SubtiList entry:'''[http://genolist.pasteur.fr/SubtiList/genome.cgi?gene_detail+BG10200]
+* '''SubtiList entry:''' [http://genolist.pasteur.fr/SubtiList/genome.cgi?gene_detail+BG10201]
 === Additional information===
@@ Line 53: / Line 54: @@
 === Basic information/ Evolution ===
-* '''Catalyzed reaction/ biological activity:''' Protein HPr N(pi)-phospho-L-histidine + protein EIIA = protein [[PtsH|HPr]] + protein EIIA N(tau)-phospho-L-histidine
+* '''Catalyzed reaction/ biological activity:''' PEP-dependent autophosphorylation on His-189, transfer of the phosphoryl group to [[PtsH |HPr]] (His-15)
-* '''Protein family:''' HPr domain HPr family
+* '''Protein family:''' PEP-utilizing enzyme family PEP-utilizing enzyme family
-* '''Paralogous protein(s):''' [[Crh]]
+* '''Paralogous protein(s):'''
 === Extended information on the protein ===
@@ Line 63: / Line 64: @@
 * '''Kinetic information:'''
-* '''Domains:''' HPr Domain (2–88)
+* '''Domains:'''
+**HPr binding site (N-Terminal Domain)
+**pyruvate binding site (C-Terminal Domain)
+**pyrophosphate/phosphate carrier histidine (central Domain)
-* '''Modification:''' phosphorylations: transient phosphorylation by [[PtsI |Enzyme I]] of the PTS on His-15, regulatory phosphorylation on Ser-46 by [[HprK]] [http://www.ncbi.nlm.nih.gov/sites/entrez/2507315 PubMed], weak phosphorylation on Ser-12 [http://www.ncbi.nlm.nih.gov/sites/entrez/17218307 PubMed], an extensive study on ''in vivo'' HPr phosphorylation can be found in Singh et al. (2008) [http://www.ncbi.nlm.nih.gov/sites/entrez/18757537  PubMed]
+* '''Modification:''' transient autophosphorylation on His-189, in vivo also phosphorylated on Ser-34 or Ser-36 [http://www.ncbi.nlm.nih.gov/pubmed/17218307 PubMed]
-* '''Cofactor(s):'''
+* '''Cofactor(s):''' Magnesium
 * '''Effectors of protein activity:'''
-* '''Interactions:''' [[GapA]]-[[PtsH|HPr]] [http://www.ncbi.nlm.nih.gov/sites/entrez/17142398 PubMed], [[PtsH|HPr]]-[[MtlR]], [[PtsH|HPr]]-[[LicR]], [[PtsH|HPr]]-[[LevR]],[[PtsH|HPr]]-[[ManR]], [[YesS]]-[[PtsH|HPr]] (HPr-His-P), [[PtsH|HPr]]-[[CcpA]] [http://www.ncbi.nlm.nih.gov/sites/entrez/12432959 PubMed], [[PtsH|HPr]]-[[RbsR]] [http://www.ncbi.nlm.nih.gov/sites/entrez/16519689 PubMed], [[HprK]]-[[PtsH|HPr]] [http://www.ncbi.nlm.nih.gov/sites/entrez/12009882 PubMed]
+* '''Interactions:'''
-* '''Localization:''' Cytoplasm  [http://www.ncbi.nlm.nih.gov/sites/entrez/16395550 PubMed]
+* '''Localization:''' Cytoplasm
 === Database entries ===
-* '''Structure:''' [http://www.rcsb.org/pdb/cgi/explore.cgi?pdbId=2HID 2HID], complex of ''L. casei'' HprK with ''B. subtilis'' HPr [http://www.ncbi.nlm.nih.gov/Structure/mmdb/mmdbsrv.cgi?Dopt=s&uid=20417 NCBI], complex of L. Casei HprK with B. Subtilis HPr-Ser-P [http://www.ncbi.nlm.nih.gov/Structure/mmdb/mmdbsrv.cgi?Dopt=s&uid=20418 NCBI]
+* '''Structure:'''
-* '''Swiss prot entry:''' [http://www.expasy.ch/cgi-bin/sprot-search-ac?P08877]
+* '''Swiss prot entry:''' [http://www.expasy.ch/cgi-bin/sprot-search-ac?P08838]
-* '''KEGG entry:''' [http://www.genome.jp/dbget-bin/www_bget?bsu+BSU13900]
+* '''KEGG entry:''' [http://www.genome.jp/dbget-bin/www_bget?bsu+BSU13910]
-* '''E.C. number:''' [http://www.expasy.org/enzyme/2.7.11.- 2.7.11.-]
+* '''E.C. number:''' [http://www.expasy.org/enzyme/2.7.3.9 2.7.3.9]
 === Additional information===
@@ Line 92: / Line 96: @@
 * '''Operon:'''
 **''[[ptsG]]-[[ptsH]]-[[ptsI]]''
-**''ptsH-[[ptsI]]''
+**''[[ptsH]]-[[ptsI]]''
 * '''[[Sigma factor]]:''' [[SigA]] [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Abstract&list_uids=+11902727 PubMed]
+* '''Regulation:''' expression activated by glucose (4.3 fold) [http://www.ncbi.nlm.nih.gov/pubmed/12850135 PubMed],  induction by glucose (''[[ptsG]]''), constitutive (''[[ptsH]]'')
-* '''Regulation:''' expression activated by glucose (2-fold) [http://www.ncbi.nlm.nih.gov/pubmed/12850135 PubMed],  induction by glucose (''[[ptsG]]''), constitutive (''[[ptsH]]'')
 * '''Regulatory mechanism:''' ''[[ptsG]]'': transcriptional antitermination via the [[GlcT]]-dependent RNA-switch
@@ Line 104: / Line 107: @@
 =Biological materials =
-* '''Mutant:''' MZ303 (cat), GP507 ptsH1 (S46A), GP506 (ptsH-H15A), available in [[Stülke]] lab
+* '''Mutant:''' GP864 (ermC), available in [[Stülke]] lab
-* '''Expression vector:''' pGP438 (with N-terminal Strep-tag, in [[pGP172]]), pAG2 (His-tag) pGP371(ptsH-S46A, with His-tag, in [[pWH844]]), available in [[Stülke]]
+* '''Expression vector:''' pAG3 (His-tag), available in [[Galinier]] lab
 * '''lacZ fusion:'''
@@ Line 112: / Line 115: @@
 * '''GFP fusion:'''
-* '''two-hybrid system:''' B. pertussis adenylate cyclase-based bacterial two hybrid system ([[BACTH]]), available in [[Boris Görke| Görke]] lab
+* '''Antibody:'''
-* '''Antibody:''' available in [[Stülke]] lab
 =Labs working on this gene/protein=
@@ Line 121: / Line 122: @@
 [[Stülke|Jörg Stülke]], University of Göttingen, Germany [http://wwwuser.gwdg.de/~genmibio/stuelke.html Homepage]
-[[Wolfgang Hillen]], Erlangen University, Germany [http://www.biologie.uni-erlangen.de/mibi/index2.html Homepage]
-[[Richard Brennan]], Houston, Texas, USA [http://www.mdanderson.org/departments/biochem/display.cfm?id=556ef368-6c81-4043-b74f350d41dd06cb&method=displayfull&pn=a8427ebd-d0ff-11d4-80fd00508b603a14 Homepage]
-[[Boris Görke]], University of Göttingen, Germany [http://wwwuser.gwdg.de/~genmibio/goerke.html Homepage]
-[[Anne Galinier]], University of Marseille, France
 =Your additional remarks=
@@ Line 135: / Line 128: @@
 # Blencke et al. (2003) Transcriptional profiling of gene expression in response to glucose in ''Bacillus subtilis'': regulation of the central metabolic pathways. ''Metab Eng.'' '''5:''' 133-149 [http://www.ncbi.nlm.nih.gov/pubmed/12850135 PubMed]
+# Frisby, D., and Zuber, P. 1994. Mutations in pts cause catabolite-resistant sporulation and altered regulation of spo0H in Bacillus subtilis. J. Bacteriol. 176: 2587-2595. [http://www.ncbi.nlm.nih.gov/sites/entrez/8169206  PubMed]
 # Macek B, Mijakovic I, Olsen JV (2007) The serine/threonine/tyrosine phosphoproteome of the model bacterium Bacillus subtilis. ''Mol Cell Proteomics'' '''6(4):''' 697-707. [http://www.ncbi.nlm.nih.gov/sites/entrez/17218307 PubMed]
-# Müller W, Horstmann N, Hillen W (2006) The transcription regulator RbsR represents a novel interaction partner of the phosphoprotein HPr-Ser46-P in Bacillus subtilis ''FEBS J.'' '''273(6):''' 1251-61. [http://www.ncbi.nlm.nih.gov/sites/entrez/16519689 PubMed]
-# 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.[http://www.ncbi.nlm.nih.gov/sites/entrez/17142398 PubMed]
-# Fieulaine, S., Morera, S., Poncet, S., Mijakovic, I., Galinier, A., Janin, J., Deutscher, J., and Nessler, S. (2002) X-ray structure of a bifunctional protein kinase in complex with its protein substrate HPr. Proc Natl Acad Sci U S A 99: 13437-13441. [http://www.ncbi.nlm.nih.gov/sites/entrez/12359875 PubMed]
-# Arnaud M, Vary P, Zagorec M, Klier A, Débarbouillé M, Postma P, Rapoport G (1992) Regulation of the sacPA operon of Bacillus subtilis: identification of phosphotransferase system components involved in SacT activity. J Bacteriol 174:3161-3170. [http://www.ncbi.nlm.nih.gov/sites/entrez/1577686 PubMed]
-# Deutscher, J., Kessler, U., Alpert, C. A., and Hengstenberg, W. (1984) Bacterial phosphoenolpyruvate-dependent phosphotransferase system: P-ser-HPr and its possible regulatory function. Biochemistry 23: 4455-4460. [http://pubs.acs.org/doi/abs/10.1021/bi00314a033 DOI:10.1021/bi00314a033]
-# Deutscher, J., Küster, E., Bergstedt, U., Charrier, V., and Hillen, W. (1995) Protein kinase-dependent HPr/CcpA interaction links glycolytic activity to carbon catabolite repression in Gram-positive bacteria. Mol. Microbiol. 15: 1049-1053. [http://www.ncbi.nlm.nih.gov/sites/entrez/7623661  PubMed]
-# Eisermann, R., Deutscher, J., Gonzy-Tréboul, G., and Hengstenberg, W. (1988) Site-directed mutagenesis with the ptsH gene of Bacillus subtilis. J Biol Chem 263: 17050-17054. [http://www.ncbi.nlm.nih.gov/sites/entrez/2846556  PubMed]
-# Frisby, D., and Zuber, P. 1994. Mutations in pts cause catabolite-resistant sporulation and altered regulation of spo0H in Bacillus subtilis. J. Bacteriol. 176: 2587-2595. [http://www.ncbi.nlm.nih.gov/sites/entrez/8169206  PubMed]
-# Galinier A, Deutscher J, Martin-Verstraete I: (1999) 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 , 286:307-314. [http://www.ncbi.nlm.nih.gov/sites/entrez/9973552  PubMed]
-# Görke, B., Fraysse, L. & Galinier, A. (2004) Drastic differences in Crh and HPr synthesis levels reflect their different impacts on catabolite repression in Bacillus subtilis. J. Bacteriol. 186, 2992-2995 . [http://www.ncbi.nlm.nih.gov/sites/entrez/15126459  PubMed]
-# Lindner, C., Galinier, A., Hecker, M. & Deutscher, J. (1999) Regulation of the activity of the Bacillus subtilis antiterminator LicT by multiple PEP-dependent, enzyme I- and HPr-catalysed phosphorylation. Mol. Microbiol. 31, 995-1006 . [http://www.ncbi.nlm.nih.gov/sites/entrez/10048041  PubMed]
-# Lindner, C., Hecker, M., Le Coq, D. & Deutscher, J. (2002) Bacillus subtilis mutant LicT antiterminators exhibiting enzyme I- and HPr-independent antitermination affect catabolite repression of the bglPH operon. J. Bacteriol. 184, 4819-4828 . [http://www.ncbi.nlm.nih.gov/sites/entrez/12169607  PubMed]
-# Martin-Verstraete, I., Charrier, V., Stülke, J., Galinier, A., Erni, B., Rapoport, G., & Deutscher, J. (1998) Antagonistic effects of dual PTS catalyzed phosphorylation on the Bacillus subtilis transcriptional activator LevR. Mol. Microbiol. 28: 293-303. [http://www.ncbi.nlm.nih.gov/sites/entrez/9622354  PubMed]
-# 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. [http://www.ncbi.nlm.nih.gov/sites/entrez/10217795  PubMed]
-# Reizer, J., Sutrina, S. L., Saier, Jr., M. H., Stewart, G. C., Peterkofsky, A., and Reddy, P. (1989) Mechanistic and physiological consequences of HPr(Ser) phosphorylation on the activities of the phosphoenolpyruvate:sugar phosphotransferase system in Gram-positive bacteria: studies with site-specific mutants of HPr. EMBO J 8: 2111-2120. [http://www.ncbi.nlm.nih.gov/sites/entrez/2507315  PubMed]
-# Schmalisch, M., Bachem, S. & Stülke, J. (2003) Control of the Bacillus subtilis antiterminator protein GlcT by phosphorylation: Elucidation of the phosphorylation chain leading to inactivation of GlcT. J. Biol. Chem. 278: 51108-51115. [http://www.ncbi.nlm.nih.gov/sites/entrez/14527945  PubMed]
-# Schumacher, M. A. et al. (2004) Structural basis for allosteric control of the transcription regulator CcpA by the phosphoprotein HPr-Ser46-P. Cell 118, 731-741 . [http://www.ncbi.nlm.nih.gov/sites/entrez/15369672  PubMed]
-# Singh, K. D., Halbedel, S., Görke, B. & Stülke, J. (2007) Control of the phosphorylation state of the HPr protein of the phosphotransferase system in Bacillus subtilis: implication of the protein phosphatase PrpC. J. Mol. Microbiol. Biotechnol. 13: 165-171. [http://www.ncbi.nlm.nih.gov/sites/entrez/17693724  PubMed]
-# Singh, K. D., Schmalisch, M. H., Stülke, J. & Görke, B. (2008) Carbon catabolite repression in Bacillus subtilis: A quantitative analysis of repression exerted by different carbon sources. J. Bacteriol. 190: 7275-7284. [http://www.ncbi.nlm.nih.gov/sites/entrez/18757537  PubMed]
-# Stülke, J., Martin-Verstraete, I., Charrier, V., Klier, A., Deutscher, J. & Rapoport, G. (1995) The HPr protein of the phosphotransferase system links induction and catabolite repression of the Bacillus subtilis levanase operon. J. Bacteriol. 177: 6928-6936. [http://www.ncbi.nlm.nih.gov/sites/entrez/7592487  PubMed]
-# Tortosa, P., Aymerich, S., Lindner, C., Saier, M.H., Jr., Reizer, J. and Le Coq, D. (1997) Multiple phosphorylation of SacY, a Bacillus subtilis antiterminator negatively controlled by the phosphotransferase system. J. Biol. Chem. 272, 17230-17237. [http://www.ncbi.nlm.nih.gov/sites/entrez/9202047  PubMed]
-# Charrier V, Buckley E, Parsonage D, Galinier A, Darbon E, Jaquinod M, Forest E, Deutscher J, Claiborne A (1997) Cloning and sequencing of two enterococcal ''glpK'' genes and regulation of the encoded glycerol kinases by phosphoenolpyruvate-dependent, phosphotransferase system-catalyzed phosphorylation of a single histidyl residue. J Biol Chem 272:14166-14174. [http://www.ncbi.nlm.nih.gov/sites/entrez/9162046 PubMed]
-# Darbon E, Servant P, Poncet S, Deutscher J (2002) Antitermination by GlpP, catabolite repression via CcpA and inducer exclusion triggered by P~GlpK dephosphorylation control ''Bacillus subtilis glpFK'' expression. Mol Microbiol 43:1039-1052. [http://www.ncbi.nlm.nih.gov/sites/entrez/11929549 PubMed]
-# Jones, B.E., Rajagopal, P., and Klevit, R.E. (1997) Phosphorylation on histidine is accompanied by localized structural changes in the phosphocarrier protein, HPr from Bacillus subtilis. Protein Sci 6: 2107-2119. [http://www.ncbi.nlm.nih.gov/sites/entrez/9336834 PubMed]
-# Rajagopal, P., Waygood, E.B., and Klevit, R.E. (1994) Structural consequences of histidine phosphorylation: NMR characterization of the phosphohistidine form of histidine-containing protein from Bacillus subtilis and Escherichia coli. Biochemistry 33: 15271-15282. [http://www.ncbi.nlm.nih.gov/sites/entrez/7803390 PubMed]

Difference between revisions of "Sandbox"

Revision as of 04:24, 12 April 2009

Contents

The gene

Basic information

Phenotypes of a mutant

Database entries

Additional information

The protein

Basic information/ Evolution

Extended information on the protein

Database entries

Additional information

Expression and regulation

Biological materials

Labs working on this gene/protein

Your additional remarks

References

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