Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
twitter

Escherichia coli K-12 substr. MG1655 Polypeptide: phosphotransferase system enzyme IIA, regulation of potassium transport



Gene: ptsN Accession Numbers: EG11682 (EcoCyc), b3204, ECK3193

Synonyms: yhbI, rpoP, EIIANtr, enzyme IIANtr

Regulation Summary Diagram: ?

Alternative forms of phosphotransferase system enzyme IIA, regulation of potassium transport: PtsN-phosphorylated

Summary:
ptsN encodes a protein paralogous to Enzyme IIAfru of the phosphoenolpyruvate (PEP)-dependent carbohydrate phosphotransferase system (PTS) [Jones94, Powell95]. PtsN regulates the activity or expression of potassium transporters [Lee07d, Luttmann09]. The PtsN protein, also known as Enzyme IIANtr, is part of a phosphorelay system (PTSNtr) in E.coli K-12 that may have a role in the regulation of nitrogen metabolism [Lee13c, PflugerGrau10] although this has been questioned [Ninfa11].

PtsN can be phosphorylated in vivo; both the energy coupling phosphotransferases of the Ntr PTS (PtsP and Npr) as well as the sugar PTS contribute to the phosphorylation of PtsN [Zimmer08]. Dephosphorylated PtsN interacts with the potassium transporter TrkA and inhibits its activity [Lee07d]. Dephosphorylated PtsN also interacts with the sensor kinase KdpD and stimulates its activity, resulting in increased levels of phosphorylated response regulator KdpE, thereby increasing expression of kdpFABC, a high-affinity potassium transporter [Luttmann09]. Via regulating the intracellular K+ concentration, PtsN affects sigma factor selectivity and thus transcription levels for a number of genes [Lee10b].

All nutrient utilization phenotypes previously attributed to lack of ptsN are synthetic phenotypes dependent on the presence of the ilvG mutation, which causes a lack of AHASII in the K-12 MG1655 and W3110 strain backgrounds [Reaves11]. The phenotypes include sensitivity of ptsN mutants to leucine-containing peptides [Lee05b] and inhibition of growth by a number of TCA cycle intermediates and carbohydrates when a nucleoside base or amino acid is used as a source of carbon, nitrogen, and energy [Powell95]. The intracellular K+ concentration has effects on expression and activity of AHASI, and thus the effect of a ptsN mutation on cellular metabolism depends on the strain background [Reaves11].

PtsN(K75D) exhibits a phosphorylation dependent mobility shift on SDS-PAGE. Glutamine inhibits and α-ketoglutarate stimulates, the in vitro phosphorylation of PtsN(K75D). This effect was also demonstrated in an ilvG+ revertant of E. coli MG1655 [Lee13c].

A Tn10 insertion in ptsN suppresses the conditionally lethal erats mutation [Powell95]. ptsN is a multicopy suppressor of σE essentiality; the ability to phosphorylate PtsN is required for suppression. Overexpression of ptsN lowers envelope stress [Hayden08].

A crystal structure of PtsN has been solved at 2.35 Å resolution [Bordo98], and the protein has been characterized in solution by NMR [Li03a, Wang05e]. While the crystal structure shows a dimer [Bordo98], PtsN is monomeric in solution; PtsN-phosphorylated is the the predominant form in LB-grown cells [Wang05e].

Comments: [Ninfa07, Ninfa11]

Review: [PflugerGrau10]

Gene Citations: [Rhodius05]

Locations: cytosol

Map Position: [3,344,600 -> 3,345,091] (72.09 centisomes)
Length: 492 bp / 163 aa

Molecular Weight of Polypeptide: 17.96 kD (from nucleotide sequence), 19.0 kD (experimental) [Jones94 ]

Unification Links: ASAP:ABE-0010520 , EchoBASE:EB1633 , EcoGene:EG11682 , EcoliWiki:b3204 , Mint:MINT-1239820 , ModBase:P69829 , OU-Microarray:b3204 , PortEco:ptsN , PR:PRO_000023634 , Pride:P69829 , Protein Model Portal:P69829 , RefSeq:NP_417671 , RegulonDB:EG11682 , SMR:P69829 , String:511145.b3204 , UniProt:P69829

Relationship Links: InterPro:IN-FAMILY:IPR002178 , InterPro:IN-FAMILY:IPR006320 , InterPro:IN-FAMILY:IPR016152 , PDB:Structure:1A6J , Pfam:IN-FAMILY:PF00359 , Prosite:IN-FAMILY:PS00372 , Prosite:IN-FAMILY:PS51094

In Paralogous Gene Group: 187 (14 members)

In Reactions of unknown directionality:

Not in pathways:
Npr phosphorylated + PtsN = PtsN-phosphorylated + Npr

GO Terms:

Biological Process: GO:0010243 - response to organonitrogen compound Inferred from experiment [Lee13c]
GO:0032412 - regulation of ion transmembrane transporter activity Inferred from experiment [Lee07d]
GO:0043086 - negative regulation of catalytic activity Inferred from experiment [Lee07d]
GO:0045860 - positive regulation of protein kinase activity Inferred from experiment [Luttmann09]
GO:0006810 - transport Inferred by computational analysis [GOA01a]
GO:0009401 - phosphoenolpyruvate-dependent sugar phosphotransferase system Inferred by computational analysis [GOA01a]
GO:0016310 - phosphorylation Inferred by computational analysis [UniProtGOA11a]
GO:0034219 - carbohydrate transmembrane transport Inferred by computational analysis [GOA01a]
Molecular Function: GO:0004857 - enzyme inhibitor activity Inferred from experiment [Lee07d]
GO:0005515 - protein binding Inferred from experiment [Rajagopala14, Luttmann09, Lee07d]
GO:0030295 - protein kinase activator activity Inferred from experiment [Luttmann09]
GO:0005215 - transporter activity Inferred by computational analysis [GOA01a]
GO:0008982 - protein-N(PI)-phosphohistidine-sugar phosphotransferase activity Inferred by computational analysis [GOA01a]
GO:0016301 - kinase activity Inferred by computational analysis [UniProtGOA11a]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005829 - cytosol Inferred from experiment Inferred by computational analysis [DiazMejia09, Ishihama08]
GO:0005737 - cytoplasm Inferred by computational analysis [UniProtGOA11, UniProtGOA11a]

MultiFun Terms: regulation type of regulation posttranscriptional inhibition / activation of enzymes

Essentiality data for ptsN knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 1]
M9 medium with 1% glycerol Yes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 2]
MOPS medium with 0.4% glucose Yes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 1]

Credits:
Last-Curated ? 02-Nov-2011 by Keseler I , SRI International


Sequence Features

Feature Class Location Common Name Citations Comment
Conserved-Region 12 -> 156  
[UniProt09]
UniProt: PTS EIIA type-2;
Phosphorylation-Modification 73 phosphorylated histidine residue
[Zimmer08]
 
Active-Site 73  
[UniProt10a]
UniProt: Tele-phosphohistidine intermediate; Non-Experimental Qualifier: by similarity;


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
10/20/97 Gene b3204 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG11682; confirmed by SwissProt match.


References

Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

Bordo98: Bordo D, van Monfort RL, Pijning T, Kalk KH, Reizer J, Saier MH, Dijkstra BW (1998). "The three-dimensional structure of the nitrogen regulatory protein IIANtr from Escherichia coli." J Mol Biol 279(1);245-55. PMID: 9636714

DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114

GOA01a: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

Hayden08: Hayden JD, Ades SE (2008). "The Extracytoplasmic Stress Factor, sigma, Is Required to Maintain Cell Envelope Integrity in Escherichia coli." PLoS ONE 3(2);e1573. PMID: 18253509

Ishihama08: Ishihama Y, Schmidt T, Rappsilber J, Mann M, Hartl FU, Kerner MJ, Frishman D (2008). "Protein abundance profiling of the Escherichia coli cytosol." BMC Genomics 9;102. PMID: 18304323

Jones94: Jones DH, Franklin FC, Thomas CM (1994). "Molecular analysis of the operon which encodes the RNA polymerase sigma factor sigma 54 of Escherichia coli." Microbiology 1994;140 ( Pt 5);1035-43. PMID: 8025669

Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394

Lee05b: Lee CR, Koo BM, Cho SH, Kim YJ, Yoon MJ, Peterkofsky A, Seok YJ (2005). "Requirement of the dephospho-form of enzyme IIANtr for derepression of Escherichia coli K-12 ilvBN expression." Mol Microbiol 58(1);334-44. PMID: 16164569

Lee07d: Lee CR, Cho SH, Yoon MJ, Peterkofsky A, Seok YJ (2007). "Escherichia coli enzyme IIANtr regulates the K+ transporter TrkA." Proc Natl Acad Sci U S A 104(10);4124-9. PMID: 17289841

Lee10b: Lee CR, Cho SH, Kim HJ, Kim M, Peterkofsky A, Seok YJ (2010). "Potassium mediates Escherichia coli enzyme IIA(Ntr) -dependent regulation of sigma factor selectivity." Mol Microbiol 78(6);1468-83. PMID: 21143318

Lee13c: Lee CR, Park YH, Kim M, Kim YR, Park S, Peterkofsky A, Seok YJ (2013). "Reciprocal regulation of the autophosphorylation of enzyme INtr by glutamine and α-ketoglutarate in Escherichia coli." Mol Microbiol 88(3);473-85. PMID: 23517463

Li03a: Li X, Peterkofsky A, Wang G (2003). "1H, 15N, and 13C chemical shift assignments of the Escherichia coli nitrogen regulatory phosphocarrier IIA(Ntr)." J Biomol NMR 27(4);401-2. PMID: 14512741

Luttmann09: Luttmann D, Heermann R, Zimmer B, Hillmann A, Rampp IS, Jung K, Gorke B (2009). "Stimulation of the potassium sensor KdpD kinase activity by interaction with the phosphotransferase protein IIA(Ntr) in Escherichia coli." Mol Microbiol 72(4);978-94. PMID: 19400808

Ninfa07: Ninfa AJ (2007). "Regulation of carbon and nitrogen metabolism: adding regulation of ion channels and another second messenger to the mix." Proc Natl Acad Sci U S A 104(11);4243-4. PMID: 17360507

Ninfa11: Ninfa AJ (2011). "Unnecessary signaling: poorly named?." J Bacteriol 193(18);4571-3. PMID: 21742895

PflugerGrau10: Pfluger-Grau K, Gorke B (2010). "Regulatory roles of the bacterial nitrogen-related phosphotransferase system." Trends Microbiol 18(5);205-14. PMID: 20202847

Powell95: Powell BS, Court DL, Inada T, Nakamura Y, Michotey V, Cui X, Reizer A, Saier MH, Reizer J (1995). "Novel proteins of the phosphotransferase system encoded within the rpoN operon of Escherichia coli. Enzyme IIANtr affects growth on organic nitrogen and the conditional lethality of an erats mutant." J Biol Chem 1995;270(9);4822-39. PMID: 7876255

Rajagopala14: Rajagopala SV, Sikorski P, Kumar A, Mosca R, Vlasblom J, Arnold R, Franca-Koh J, Pakala SB, Phanse S, Ceol A, Hauser R, Siszler G, Wuchty S, Emili A, Babu M, Aloy P, Pieper R, Uetz P (2014). "The binary protein-protein interaction landscape of Escherichia coli." Nat Biotechnol 32(3);285-90. PMID: 24561554

Reaves11: Reaves ML, Rabinowitz JD (2011). "Characteristic phenotypes associated with ptsN-null mutants in Escherichia coli K-12 are absent in strains with functional ilvG." J Bacteriol 193(18);4576-81. PMID: 21531803

Rhodius05: Rhodius VA, Suh WC, Nonaka G, West J, Gross CA (2005). "Conserved and variable functions of the sigmaE stress response in related genomes." PLoS Biol 4(1);e2. PMID: 16336047

UniProt09: UniProt Consortium (2009). "UniProt version 15.8 released on 2009-10-01 00:00:00." Database.

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on the manual assignment of UniProtKB Subcellular Location terms in UniProtKB/Swiss-Prot entries."

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

Wang05e: Wang G, Peterkofsky A, Keifer PA, Li X (2005). "NMR characterization of the Escherichia coli nitrogen regulatory protein IIANtr in solution and interaction with its partner protein, NPr." Protein Sci 14(4);1082-90. PMID: 15741344

Zimmer08: Zimmer B, Hillmann A, Gorke B (2008). "Requirements for the phosphorylation of the Escherichia coli EIIANtr protein in vivo." FEMS Microbiol Lett 286(1);96-102. PMID: 18625021

Other References Related to Gene Regulation

Kumar11: Kumar R, Shimizu K (2011). "Transcriptional regulation of main metabolic pathways of cyoA, cydB, fnr, and fur gene knockout Escherichia coli in C-limited and N-limited aerobic continuous cultures." Microb Cell Fact 10;3. PMID: 21272324


Report Errors or Provide Feedback
Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Mon Nov 24, 2014, BIOCYC14A.