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Escherichia coli K-12 substr. MG1655 Transporter: N-acetylneuraminate:H+ symporter



Gene: nanT Accession Numbers: G436 (EcoCyc), b3224, ECK3213

Regulation Summary Diagram: ?

Summary:
NanT is a sialic acid:H+ symporter in E. coli K-12.

Purified NanT, reconstituted into liposomes, functions as a bidirectional secondary transporter in vitro [Mulligan12]. NanT facilitates both substrate counterflow and substrate exchange in vitro [Mulligan09, Mulligan12]. NanT transport is energised by the proton gradient alone [Mulligan12]. nanT expressed from a plasmid, restores sialic acid transport and utilisation in mutants unable to transport sialic acid [Martinez95]. Whole cell transport experiments in E.coli strain K235 indicate that sialic acid is transported with an affinity of 30 μM and uptake is non-competitively inhibited by N-acetylglucosamine [RodriguezAparic87]. Sialic acid transport is fully disrupted by 2,4-dinitrophenol, but only partially by sodium arsenate, suggesting that NanT is a sugar/proton symporter [RodriguezAparic87].

NanT is a member of the major facilitator superfamily and has fourteen predicted transmembrane segments with the C-terminus and N-terminus locted in the cytosol [Martinez95]. Expression of nanT is induced by sialic acid [Vimr85]. Imported sialic acid is cleaved by NanA to yield N-acetylmannosamine and pyruvate.

Regulation has been described [Kalivoda03]. Transcription of nanATEK-yhcH (sialic acid catabolic operon) is repressed by a NanR homodimer [Kalivoda03].

Gene Citations: [Ohta85, Plumbridge99, Kalivoda13]

Locations: inner membrane

Map Position: [3,369,106 <- 3,370,596] (72.62 centisomes)
Length: 1491 bp / 496 aa

Molecular Weight of Polypeptide: 53.551 kD (from nucleotide sequence)

Unification Links: ASAP:ABE-0010577 , CGSC:18121 , EchoBASE:EB4145 , EcoGene:EG20145 , EcoliWiki:b3224 , ModBase:P41036 , OU-Microarray:b3224 , PortEco:nanT , Protein Model Portal:P41036 , RefSeq:NP_417691 , RegulonDB:G436 , SMR:P41036 , String:511145.b3224 , UniProt:P41036

Relationship Links: InterPro:IN-FAMILY:IPR004742 , InterPro:IN-FAMILY:IPR005828 , InterPro:IN-FAMILY:IPR011701 , InterPro:IN-FAMILY:IPR016196 , InterPro:IN-FAMILY:IPR020846 , Pfam:IN-FAMILY:PF00083 , Pfam:IN-FAMILY:PF07690 , Prosite:IN-FAMILY:PS00216 , Prosite:IN-FAMILY:PS00217 , Prosite:IN-FAMILY:PS50850

In Paralogous Gene Group: 136 (7 members)

Gene-Reaction Schematic: ?

Instance reaction of [N-acetylneuraminate[periplasmic space] + H+[periplasmic space]N-acetylneuraminate[cytosol] + H+[cytosol]] (no EC#):
i1: N-acetyl-β-neuraminate[periplasmic space] + H+[periplasmic space]N-acetyl-β-neuraminate[cytosol] + H+[cytosol] (no EC#)

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0015739 - sialic acid transport Inferred from experiment Inferred by computational analysis [GOA06, Mulligan12, Vimr85]
GO:0015992 - proton transport Inferred by computational analysis Inferred from experiment [Mulligan12, GOA06]
GO:0006810 - transport Inferred by computational analysis [UniProtGOA11a]
GO:0008643 - carbohydrate transport Inferred by computational analysis [UniProtGOA11a]
GO:0034219 - carbohydrate transmembrane transport Inferred by computational analysis [GOA01a]
GO:0055085 - transmembrane transport Inferred by computational analysis [GOA01a, Martinez95]
Molecular Function: GO:0005351 - sugar:proton symporter activity Inferred from experiment [Mulligan12]
GO:0015136 - sialic acid transmembrane transporter activity Inferred from experiment [Vimr85, Mulligan12]
GO:0015144 - carbohydrate transmembrane transporter activity Inferred by computational analysis [GOA01a, Martinez95]
GO:0015538 - sialic acid:proton symporter activity Inferred by computational analysis [GOA06]
GO:0022857 - transmembrane transporter activity Inferred by computational analysis [GOA01a]
Cellular Component: GO:0005886 - plasma membrane Inferred from experiment Inferred by computational analysis [UniProtGOA11, UniProtGOA11a, GOA06, DiazMejia09, Daley05]
GO:0005887 - integral component of plasma membrane Inferred by computational analysis [Martinez95]
GO:0016020 - membrane Inferred by computational analysis [UniProtGOA11a]
GO:0016021 - integral component of membrane Inferred by computational analysis [UniProtGOA11a, GOA01a]

MultiFun Terms: cell structure membrane
metabolism central intermediary metabolism amino sugar conversions
transport Electrochemical potential driven transporters Porters (Uni-, Sym- and Antiporters)

Essentiality data for nanT knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 1% glycerol Yes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucose Yes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
Yes [Feist07, Comment 4]

Enzymatic reaction of: N-acetylneuraminate:H+ symporter


Enzymatic reaction of: N-acetyl-β-neuraminate:H+ symporter (N-acetylneuraminate:H+ symporter)

Credits:
Imported from MetaCyc 04-Feb-2014 by Caspi R , SRI International


Sequence Features

Feature Class Location Citations Comment
Transmembrane-Region 22 -> 42
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 58 -> 78
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 92 -> 112
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 148 -> 168
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 170 -> 190
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Sequence-Conflict 221
[Martinez95, UniProt10]
Alternate sequence: E → A; UniProt: (in Ref. 1; AAA86827);
Transmembrane-Region 224 -> 244
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 247 -> 267
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 278 -> 298
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 313 -> 333
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 355 -> 375
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 406 -> 426
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 431 -> 451
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Sequence-Conflict 441
[Martinez95, UniProt10]
Alternate sequence: S → R; UniProt: (in Ref. 1; AAA86827);


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
Peter D. Karp on Wed Jan 18, 2006:
Gene right-end position adjusted based on analysis performed in the 2005 E. coli annotation update [Riley06 ].
10/20/97 Gene b3224 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene G436.


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

Daley05: Daley DO, Rapp M, Granseth E, Melen K, Drew D, von Heijne G (2005). "Global topology analysis of the Escherichia coli inner membrane proteome." Science 308(5726);1321-3. PMID: 15919996

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

Feist07: Feist AM, Henry CS, Reed JL, Krummenacker M, Joyce AR, Karp PD, Broadbelt LJ, Hatzimanikatis V, Palsson BO (2007). "A genome-scale metabolic reconstruction for Escherichia coli K-12 MG1655 that accounts for 1260 ORFs and thermodynamic information." Mol Syst Biol 3;121. PMID: 17593909

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

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

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

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

Kalivoda03: Kalivoda KA, Steenbergen SM, Vimr ER, Plumbridge J (2003). "Regulation of sialic acid catabolism by the DNA binding protein NanR in Escherichia coli." J Bacteriol 185(16);4806-15. PMID: 12897000

Kalivoda13: Kalivoda KA, Steenbergen SM, Vimr ER (2013). "Control of the Escherichia coli sialoregulon by transcriptional repressor NanR." J Bacteriol 195(20);4689-701. PMID: 23935044

Martinez95: Martinez J, Steenbergen S, Vimr E (1995). "Derived structure of the putative sialic acid transporter from Escherichia coli predicts a novel sugar permease domain." J Bacteriol 1995;177(20);6005-10. PMID: 7592358

Mulligan09: Mulligan C, Geertsma ER, Severi E, Kelly DJ, Poolman B, Thomas GH (2009). "The substrate-binding protein imposes directionality on an electrochemical sodium gradient-driven TRAP transporter." Proc Natl Acad Sci U S A 106(6);1778-83. PMID: 19179287

Mulligan12: Mulligan C, Leech AP, Kelly DJ, Thomas GH (2012). "The membrane proteins SiaQ and SiaM form an essential stoichiometric complex in the sialic acid tripartite ATP-independent periplasmic (TRAP) transporter SiaPQM (VC1777-1779) from Vibrio cholerae." J Biol Chem 287(5);3598-608. PMID: 22167185

Ohta85: Ohta Y, Watanabe K, Kimura A (1985). "Complete nucleotide sequence of the E. coli N-acetylneuraminate lyase." Nucleic Acids Res 13(24);8843-52. PMID: 3909108

Plumbridge99: Plumbridge J, Vimr E (1999). "Convergent pathways for utilization of the amino sugars N-acetylglucosamine, N-acetylmannosamine, and N-acetylneuraminic acid by Escherichia coli." J Bacteriol 1999;181(1);47-54. PMID: 9864311

Riley06: Riley M, Abe T, Arnaud MB, Berlyn MK, Blattner FR, Chaudhuri RR, Glasner JD, Horiuchi T, Keseler IM, Kosuge T, Mori H, Perna NT, Plunkett G, Rudd KE, Serres MH, Thomas GH, Thomson NR, Wishart D, Wanner BL (2006). "Escherichia coli K-12: a cooperatively developed annotation snapshot--2005." Nucleic Acids Res 34(1);1-9. PMID: 16397293

RodriguezAparic87: Rodriguez-Aparicio LB, Reglero A, Luengo JM (1987). "Uptake of N-acetylneuraminic acid by Escherichia coli K-235. Biochemical characterization of the transport system." Biochem J 1987;246(2);287-94. PMID: 2825630

UniProt10: UniProt Consortium (2010). "UniProt version 2010-11 released on 2010-11-02 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."

Vimr85: Vimr ER, Troy FA (1985). "Identification of an inducible catabolic system for sialic acids (nan) in Escherichia coli." J Bacteriol 164(2);845-53. PMID: 3902799

Other References Related to Gene Regulation

Beisel11: Beisel CL, Storz G (2011). "The base-pairing RNA spot 42 participates in a multioutput feedforward loop to help enact catabolite repression in Escherichia coli." Mol Cell 41(3);286-97. PMID: 21292161

Beisel12: Beisel CL, Updegrove TB, Janson BJ, Storz G (2012). "Multiple factors dictate target selection by Hfq-binding small RNAs." EMBO J 31(8);1961-74. PMID: 22388518

Bradley07: Bradley MD, Beach MB, de Koning AP, Pratt TS, Osuna R (2007). "Effects of Fis on Escherichia coli gene expression during different growth stages." Microbiology 153(Pt 9);2922-40. PMID: 17768236


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 Thu Dec 18, 2014, BIOCYC13A.