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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) [Jones94a, Powell95]. PtsN regulates the activity or expression of potassium transporters [Lee07, 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 [Lee13b, 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 [Lee07]. 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 [Lee10a].

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 [Lee13b].

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, Wang05b]. While the crystal structure shows a dimer [Bordo98], PtsN is monomeric in solution; PtsN-phosphorylated is the the predominant form in LB-grown cells [Wang05b].

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) [Jones94a ]

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 [Lee13b]
GO:0032412 - regulation of ion transmembrane transporter activity Inferred from experiment [Lee07]
GO:0043086 - negative regulation of catalytic activity Inferred from experiment [Lee07]
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 [Lee07]
GO:0005515 - protein binding Inferred from experiment [Rajagopala14, Luttmann09, Lee07]
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

Jones94a: 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

Lee07: 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

Lee10a: 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

Lee13b: 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."

Wang05b: 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


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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 Fri Dec 19, 2014, BIOCYC14A.