Escherichia coli K-12 substr. MG1655 Enzyme: 4-amino-4-deoxy-L-arabinose (L-Ara4N) transferase

Gene: arnT Accession Numbers: G7170 (EcoCyc), b2257, ECK2250

Synonyms: yfbI, pmrK

Regulation Summary Diagram: ?

Regulation summary diagram for arnT

ArnT is a transferase that catalyzes the addition of L-Ara4N residues from 4-amino-4-deoxy-α-L-arabinopyranosyl ditrans,octacis-undecaprenyl phosphate to lipid A. This modification confers on the organism resistance to the antibiotic polymyxin [Trent01, Trent01a]. It should be noted that K-12 derived strains express ArnT, but do not synthesize 4-amino-4-deoxy-α-L-arabinopyranosyl ditrans,octacis-undecaprenyl phosphate under normal conditions, and thus are sensitive to polymyxin.

The enzyme from E. coli has not been studied much, but the Salmonella typhimurium enzyme has been purified and characterized [Trent01, Bretscher06].

It has been suggested, although not proven, that the enzyme is involved not only in the attachment of L-Ara4N to lipid A but also in the transport of L-Ara4N from the cytoplasm, where it is synthesized, to the periplasmic side of the inner membrane, where the attachment takes place.

Deletion of arnT results in defective attachment of L-Ara4N to the lipid A core, but does not affect the transport of undecaprenyl phosphate-α-L-Ara4N across the inner membrane [Yan07].

A lpxM mutant lacking the secondary linked myristoyl group of lipid A is unable to attach L-Ara4N to lipid A [Tran05].

Expression of the arnBCADTEF operon increased during growth with elevated FeSO4, FeCl3, or ZnSO4 and was dependent upon the BasSR two-component signal transduction system [Hagiwara04, Lee05d].

Citations: [Zhou99a, Nummila95, Groisman01, Gibbons05]

Locations: inner membrane

Map Position: [2,368,930 -> 2,370,582] (51.06 centisomes, 184°)
Length: 1653 bp / 550 aa

Molecular Weight of Polypeptide: 62.543 kD (from nucleotide sequence)

Unification Links: ASAP:ABE-0007464 , EchoBASE:EB3846 , EcoGene:EG14093 , EcoliWiki:b2257 , OU-Microarray:b2257 , PortEco:arnT , PR:PRO_000022141 , Pride:P76473 , Protein Model Portal:P76473 , RefSeq:NP_416760 , RegulonDB:G7170 , String:511145.b2257 , UniProt:P76473

Relationship Links: CAZy:IN-FAMILY:GT83 , InterPro:IN-FAMILY:IPR003342 , InterPro:IN-FAMILY:IPR022839 , Pfam:IN-FAMILY:PF02366

In Paralogous Gene Group: 4 (3 members)

Gene-Reaction Schematic: ?

Gene-Reaction Schematic

Genetic Regulation Schematic: ?

Genetic regulation schematic for arnT

GO Terms:

Biological Process: GO:0009103 - lipopolysaccharide biosynthetic process Inferred from experiment Inferred by computational analysis [UniProtGOA11a, Trent01a]
GO:0010041 - response to iron(III) ion Inferred from experiment [Chamnongpol02, Hagiwara04]
GO:0006493 - protein O-linked glycosylation Inferred by computational analysis [GOA01a]
GO:0006629 - lipid metabolic process Inferred by computational analysis [UniProtGOA11a]
GO:0009245 - lipid A biosynthetic process Inferred by computational analysis [UniProtGOA11a, GOA06]
GO:0097502 - mannosylation Inferred by computational analysis [GOA01a]
Molecular Function: GO:0016763 - transferase activity, transferring pentosyl groups Inferred from experiment Inferred by computational analysis [GOA01a, Yan07]
GO:0000030 - mannosyltransferase activity Inferred by computational analysis [GOA01a]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11a, GOA06]
GO:0016757 - transferase activity, transferring glycosyl groups Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005886 - plasma membrane Inferred from experiment Inferred by computational analysis [UniProtGOA11, UniProtGOA11a, GOA06, DiazMejia09, Daley05]
GO:0016020 - membrane Inferred by computational analysis [UniProtGOA11a, GOA01a]
GO:0016021 - integral component of membrane Inferred by computational analysis [UniProtGOA11a]

MultiFun Terms: metabolism biosynthesis of macromolecules (cellular constituents) lipopolysaccharide lipid A

Essentiality data for arnT 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]

Curated 01-Aug-2008 by Johnson A , JCVI
Last-Curated ? 10-Mar-2011 by Caspi R , SRI International

Enzymatic reaction of: 4-amino-4-deoxy-L-arabinose transferase (4-amino-4-deoxy-L-arabinose (L-Ara4N) transferase)

Synonyms: 4-amino-4-deoxy-L-arabinose transferase, L-Ara4N transferase

EC Number:

α-D-Kdo-(2→4)-α-D-Kdo-(2→6)-lipid A[periplasmic space] + 2 4-amino-4-deoxy-α-L-arabinopyranosyl ditrans,octacis-undecaprenyl phosphate[periplasmic space] <=> 4'-α-L-Ara4N-α-Kdo-(2→4)-α-Kdo-(2→6)-[P4'-α-L-ara4N]-lipid A[periplasmic space] + 2 di-trans,octa-cis-undecaprenyl phosphate[periplasmic space]

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction of enzyme catalysis.

The reaction is physiologically favored in the direction shown.

In Pathways: polymyxin resistance

Sequence Features

Protein sequence of 4-amino-4-deoxy-L-arabinose (L-Ara4N) transferase with features indicated

Feature Class Location Citations Comment
Transmembrane-Region 7 -> 27
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 81 -> 101
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 110 -> 131
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 137 -> 154
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 165 -> 185
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 204 -> 224
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 255 -> 275
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 288 -> 308
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 315 -> 335
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 346 -> 366
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 383 -> 403
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 406 -> 426
UniProt: Helical;; Non-Experimental Qualifier: potential;

Gene Local Context (not to scale): ?

Gene local context diagram

Transcription Unit:

Transcription-unit diagram


Markus Krummenacker on Tue Oct 14, 1997:
Gene object created from Blattner lab Genbank (v. M52) entry.


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

Bretscher06: Bretscher LE, Morrell MT, Funk AL, Klug CS (2006). "Purification and characterization of the L-Ara4N transferase protein ArnT from Salmonella typhimurium." Protein Expr Purif 46(1);33-9. PMID: 16226890

Chamnongpol02: Chamnongpol S, Dodson W, Cromie MJ, Harris ZL, Groisman EA (2002). "Fe(III)-mediated cellular toxicity." Mol Microbiol 45(3);711-9. PMID: 12139617

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

Gibbons05: Gibbons HS, Kalb SR, Cotter RJ, Raetz CR (2005). "Role of Mg2+ and pH in the modification of Salmonella lipid A after endocytosis by macrophage tumour cells." Mol Microbiol 55(2);425-40. PMID: 15659161

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."

Groisman01: Groisman EA (2001). "The pleiotropic two-component regulatory system PhoP-PhoQ." J Bacteriol 183(6);1835-42. PMID: 11222580

Hagiwara04: Hagiwara D, Yamashino T, Mizuno T (2004). "A Genome-wide view of the Escherichia coli BasS-BasR two-component system implicated in iron-responses." Biosci Biotechnol Biochem 68(8);1758-67. PMID: 15322361

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

Lee05d: Lee LJ, Barrett JA, Poole RK (2005). "Genome-wide transcriptional response of chemostat-cultured Escherichia coli to zinc." J Bacteriol 187(3);1124-34. PMID: 15659689

Nummila95: Nummila K, Kilpelainen I, Zahringer U, Vaara M, Helander IM (1995). "Lipopolysaccharides of polymyxin B-resistant mutants of Escherichia coli are extensively substituted by 2-aminoethyl pyrophosphate and contain aminoarabinose in lipid A." Mol Microbiol 16(2);271-8. PMID: 7565089

Tran05: Tran AX, Lester ME, Stead CM, Raetz CR, Maskell DJ, McGrath SC, Cotter RJ, Trent MS (2005). "Resistance to the antimicrobial peptide polymyxin requires myristoylation of Escherichia coli and Salmonella typhimurium lipid A." J Biol Chem 280(31);28186-94. PMID: 15951433

Trent01: Trent MS, Ribeiro AA, Lin S, Cotter RJ, Raetz CR (2001). "An inner membrane enzyme in Salmonella and Escherichia coli that transfers 4-amino-4-deoxy-L-arabinose to lipid A: induction on polymyxin-resistant mutants and role of a novel lipid-linked donor." J Biol Chem 276(46);43122-31. PMID: 11535604

Trent01a: Trent MS, Ribeiro AA, Doerrler WT, Lin S, Cotter RJ, Raetz CR (2001). "Accumulation of a polyisoprene-linked amino sugar in polymyxin-resistant Salmonella typhimurium and Escherichia coli: structural characterization and transfer to lipid A in the periplasm." J Biol Chem 276(46);43132-44. PMID: 11535605

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."

Yan07: Yan A, Guan Z, Raetz CR (2007). "An undecaprenyl phosphate-aminoarabinose flippase required for polymyxin resistance in Escherichia coli." J Biol Chem 282(49);36077-89. PMID: 17928292

Zhou99a: Zhou Z, Lin S, Cotter RJ, Raetz CR (1999). "Lipid A modifications characteristic of Salmonella typhimurium are induced by NH4VO3 in Escherichia coli K12. Detection of 4-amino-4-deoxy-L-arabinose, phosphoethanolamine and palmitate." J Biol Chem 274(26);18503-14. PMID: 10373459

Other References Related to Gene Regulation

Froelich06: Froelich JM, Tran K, Wall D (2006). "A pmrA constitutive mutant sensitizes Escherichia coli to deoxycholic acid." J Bacteriol 188(3);1180-3. PMID: 16428424

Ogasawara12: Ogasawara H, Shinohara S, Yamamoto K, Ishihama A (2012). "Novel regulation targets of the metal-response BasS-BasR two-component system of Escherichia coli." Microbiology 158(Pt 6);1482-92. PMID: 22442305

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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 19.0 on Tue Aug 4, 2015, biocyc14.