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Escherichia coli K-12 substr. MG1655 Enzyme: 10-formyltetrahydrofolate:L-methionyl-tRNAfMet N-formyltransferase



Gene: fmt Accession Numbers: EG11268 (EcoCyc), b3288, ECK3274

Synonyms: ftm, yhdD

Regulation Summary Diagram: ?

Summary:
10-formyltetrahydrofolate:L-methionyl-tRNAfMet N-formyltransferase (Fmt) attaches a formyl group to the free amino group of methionyl-tRNAfMet [Kahn80, Guillon92]. There are two types of methionine-specific tRNAs in E. coli; the initiator tRNAmet is used in the initiation of translation, while a cytidine34 in tRNAmet is used during peptide chain elongation. The formylation of the methionine-charged initiator tRNA that is catalyzed by this enzyme is important in restricting the use of the methionylated initiator tRNA specifically for initiation, rather than chain elongation [Guillon93].

Substrate recognition determinants in Fmt [Schmitt96, Gite97, Ramesh97, Ramesh98, Schmitt98, Ramesh99, Li00] and specificity determinants within the tRNAfMet [Lee91, Guillon92a, Lee92, Dyson93, Lee93, Wallis95, Li96c, Zuleeg00, Mayer02] have been studied. Upon interaction with the acceptor stem of tRNAfMet, a previously unstructured insertion loop in the N-terminal domain of Fmt undergoes a structural change by induced fit [Ramesh99]. Overexpression of Fmt can rescue the activity of tRNA mutants that are normally poor substrates for Fmt [Mangroo95, Mayer03]. D-amino acid-charged tRNAfMet can act as a substrate for Fmt at low efficiency [Goto08].

Lysine residues in the linker region are important for enzymatic activity of Fmt, while the C-terminal domain is required for non-specific binding of tRNA [Gite00].

Fmt is monomeric [Kahn80, Schmitt96a]. Crystal structures of the enzyme alone [Schmitt96] and in a complex with formyl-methionyl-tRNAfMet [Schmitt98] have been solved at 2.0 Å and 2.8 Å resolution, respectively. An N-terminal Rossmann-fold domain is linked to a C-terminal domain resembling an OB fold which is able to bind tRNAfMet [Schmitt96].

fmt appears to be constitutively expressed [Meinnel93]. fmt mutations result in a growth defect and heat sensitivity [Guillon92, Newton99].

Reviews: [Meinnel93a, RajBhandary94, Mangroo95a, Schmitt96b, Vaughan02]

Citations: [Lennette70]

Gene Citations: [Mazel94]

Locations: cytosol

Map Position: [3,432,236 -> 3,433,183] (73.98 centisomes)
Length: 948 bp / 315 aa

Molecular Weight of Polypeptide: 34.168 kD (from nucleotide sequence), 32.0 kD (experimental) [Kahn80 ]

Unification Links: ASAP:ABE-0010781 , CGSC:33616 , DIP:DIP-9668N , EchoBASE:EB1247 , EcoGene:EG11268 , EcoliWiki:b3288 , ModBase:P23882 , OU-Microarray:b3288 , PortEco:fmt , PR:PRO_000022675 , Pride:P23882 , Protein Model Portal:P23882 , RefSeq:NP_417746 , RegulonDB:EG11268 , SMR:P23882 , String:511145.b3288 , UniProt:P23882

Relationship Links: InterPro:IN-FAMILY:IPR001555 , InterPro:IN-FAMILY:IPR002376 , InterPro:IN-FAMILY:IPR005793 , InterPro:IN-FAMILY:IPR005794 , InterPro:IN-FAMILY:IPR011034 , InterPro:IN-FAMILY:IPR015518 , Panther:IN-FAMILY:PTHR11138 , PDB:Structure:1FMT , PDB:Structure:2FMT , Pfam:IN-FAMILY:PF00551 , Pfam:IN-FAMILY:PF02911 , Prosite:IN-FAMILY:PS00373

In Paralogous Gene Group: 397 (3 members)

Gene-Reaction Schematic: ?

GO Terms:

Biological Process: GO:0006413 - translational initiation Inferred by computational analysis Inferred from experiment [Kahn80, GOA06, GOA01a, GOA01]
GO:0019988 - charged-tRNA amino acid modification Inferred from experiment [Kahn80]
GO:0006412 - translation Inferred by computational analysis [UniProtGOA11, GOA06]
GO:0009058 - biosynthetic process Inferred by computational analysis [GOA01]
GO:0032259 - methylation Inferred by computational analysis [GOA01]
GO:0071951 - conversion of methionyl-tRNA to N-formyl-methionyl-tRNA Inferred by computational analysis [GOA01]
Molecular Function: GO:0004479 - methionyl-tRNA formyltransferase activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01a, GOA01, Kahn80]
GO:0003824 - catalytic activity Inferred by computational analysis [GOA01]
GO:0008168 - methyltransferase activity Inferred by computational analysis [GOA01]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11]
GO:0016742 - hydroxymethyl-, formyl- and related transferase activity Inferred by computational analysis [GOA01]
Cellular Component: GO:0005829 - cytosol Inferred from experiment [Ishihama08, LopezCampistrou05]
GO:0005737 - cytoplasm

MultiFun Terms: information transfer RNA related tRNA

Essentiality data for fmt knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB Lennox No 37 Aerobic 7   No [Baba06, Comment 1]

Credits:
Last-Curated ? 03-Dec-2009 by Keseler I , SRI International


Enzymatic reaction of: 10-formyltetrahydrofolate:L-methionyl-tRNAfMet N-formyltransferase

Synonyms: MTF, methionyl-tRNA transformylase

EC Number: 2.1.2.9

an N10-formyl-tetrahydrofolate + L-methionyl-tRNAfmet <=> a tetrahydrofolate + N-formyl-L-methionyl-tRNAfmet

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.

Summary:
Partial purification of the enzyme was first reported from E. coli B [Dickerman67].

The enzyme is specific for mehtionyl-initiator tRNAmet; it does not formylate methionyl-a cytidine34 in tRNAmet. The enzyme is able to bind uncharged tRNAfMet, but it has higher affinity for the charged methionyl-tRNAfMet [Kahn80].

Cofactors or Prosthetic Groups: Mg2+ [Kahn80], K+ [Kahn80]

Kinetic Parameters:

Substrate
Km (μM)
Citations
L-methionyl-tRNAfmet
0.35
[Kahn80]
an N10-formyl-tetrahydrofolate
13.3
[Dickerman67, BRENDA14]
an N10-formyl-tetrahydrofolate
13.5
[Kahn80]

pH(opt): 7.5 [BRENDA14, Dickerman67]


Sequence Features

Feature Class Location Common Name Citations Comment
Cleavage-of-Initial-Methionine 1  
[UniProt10a]
UniProt: Removed;
Protein-Segment 2 -> 189  
[UniProt10]
UniProt: N-terminal domain; Sequence Annotation Type: region of interest;
Chain 2 -> 315  
[UniProt09]
UniProt: Methionyl-tRNA formyltransferase;
Amino-Acid-Sites-That-Bind 43 initiator tRNA interaction determinant
[Ramesh98]
 
Protein-Segment 113 -> 116  
[UniProt10]
UniProt: Tetrahydrofolate (THF) binding; Sequence Annotation Type: region of interest;
Protein-Segment 210 -> 315  
[UniProt10]
UniProt: C-terminal domain; Sequence Annotation Type: region of interest;


Gene Local Context (not to scale): ?

Transcription Unit:

Notes:

History:
10/20/97 Gene b3288 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG11268; 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

BRENDA14: BRENDA team (2014). "Imported from BRENDA version existing on Aug 2014." http://www.brenda-enzymes.org.

Dickerman67: Dickerman HW, Steers E, Redfield BG, Weissbach H (1967). "Methionyl soluble ribonucleic acid transformylase. I. Purification and partial characterization." J Biol Chem 242(7);1522-5. PMID: 5337045

Dyson93: Dyson MR, Mandal N, RajBhandary UL (1993). "Relationship between the structure and function of Escherichia coli initiator tRNA." Biochimie 75(12);1051-60. PMID: 7515283

Gite00: Gite S, Li Y, Ramesh V, RajBhandary UL (2000). "Escherichia coli methionyl-tRNA formyltransferase: role of amino acids conserved in the linker region and in the C-terminal domain on the specific recognition of the initiator tRNA." Biochemistry 39(9);2218-26. PMID: 10694387

Gite97: Gite S, RajBhandary UL (1997). "Lysine 207 as the site of cross-linking between the 3'-end of Escherichia coli initiator tRNA and methionyl-tRNA formyltransferase." J Biol Chem 272(8);5305-12. PMID: 9030604

GOA01: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA01a: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

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

Goto08: Goto Y, Murakami H, Suga H (2008). "Initiating translation with D-amino acids." RNA 14(7);1390-8. PMID: 18515548

Guillon92: Guillon JM, Mechulam Y, Schmitter JM, Blanquet S, Fayat G (1992). "Disruption of the gene for Met-tRNA(fMet) formyltransferase severely impairs growth of Escherichia coli." J Bacteriol 174(13);4294-301. PMID: 1624424

Guillon92a: Guillon JM, Meinnel T, Mechulam Y, Lazennec C, Blanquet S, Fayat G (1992). "Nucleotides of tRNA governing the specificity of Escherichia coli methionyl-tRNA(fMet) formyltransferase." J Mol Biol 224(2);359-67. PMID: 1373194

Guillon93: Guillon JM, Mechulam Y, Blanquet S, Fayat G (1993). "Importance of formylability and anticodon stem sequence to give a tRNA(Met) an initiator identity in Escherichia coli." J Bacteriol 175(14);4507-14. PMID: 8331078

Hountondji80: Hountondji C, Fayat G, Blanquet S (1980). "Transfer RNA labeling of Escherichia coli methionyl-tRNA transformylase." Eur J Biochem 107(2);403-7. PMID: 6995120

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

Kahn80: Kahn D, Fromant M, Fayat G, Dessen P, Blanquet S (1980). "Methionyl-transfer-RNA transformylase from Escherichia coli. Purification and characterisation." Eur J Biochem 105(3);489-97. PMID: 6989606

Lee91: Lee CP, Seong BL, RajBhandary UL (1991). "Structural and sequence elements important for recognition of Escherichia coli formylmethionine tRNA by methionyl-tRNA transformylase are clustered in the acceptor stem." J Biol Chem 266(27);18012-7. PMID: 1917939

Lee92: Lee CP, Dyson MR, Mandal N, Varshney U, Bahramian B, RajBhandary UL (1992). "Striking effects of coupling mutations in the acceptor stem on recognition of tRNAs by Escherichia coli Met-tRNA synthetase and Met-tRNA transformylase." Proc Natl Acad Sci U S A 89(19);9262-6. PMID: 1409632

Lee93: Lee CP, Mandal N, Dyson MR, RajBhandary UL (1993). "The discriminator base influences tRNA structure at the end of the acceptor stem and possibly its interaction with proteins." Proc Natl Acad Sci U S A 90(15);7149-52. PMID: 8346229

Lennette70: Lennette ET, Apiriaon D (1970). "The level of Fmt-tRNA on ribosomes from streptomycin treated cells." Biochem Biophys Res Commun 41(3);804-11. PMID: 4920879

Li00: Li Y, Ramesh V, Mangroo D, Taneja C, RajBhandary UL (2000). "Suppressor mutations in Escherichia coli methionyl-tRNA formyltransferase that compensate for the formylation defect of a mutant tRNA aminoacylated with lysine." Biochemistry 39(27);8039-46. PMID: 10891086

Li96c: Li S, Kumar NV, Varshney U, RajBhandary UL (1996). "Important role of the amino acid attached to tRNA in formylation and in initiation of protein synthesis in Escherichia coli." J Biol Chem 271(2);1022-8. PMID: 8557626

LopezCampistrou05: Lopez-Campistrous A, Semchuk P, Burke L, Palmer-Stone T, Brokx SJ, Broderick G, Bottorff D, Bolch S, Weiner JH, Ellison MJ (2005). "Localization, annotation, and comparison of the Escherichia coli K-12 proteome under two states of growth." Mol Cell Proteomics 4(8);1205-9. PMID: 15911532

Mangroo95: Mangroo D, RajBhandary UL (1995). "Mutants of Escherichia coli initiator tRNA defective in initiation. Effects of overproduction of methionyl-tRNA transformylase and the initiation factors IF2 and IF3." J Biol Chem 270(20);12203-9. PMID: 7538134

Mangroo95a: Mangroo D, Wu XQ, RajBhandary UL (1995). "Escherichia coli initiator tRNA: structure-function relationships and interactions with the translational machinery." Biochem Cell Biol 73(11-12);1023-31. PMID: 8722017

Mayer02: Mayer C, RajBhandary UL (2002). "Conformational change of Escherichia coli initiator methionyl-tRNA(fMet) upon binding to methionyl-tRNA formyl transferase." Nucleic Acids Res 30(13);2844-50. PMID: 12087168

Mayer03: Mayer C, Kohrer C, Kenny E, Prusko C, RajBhandary UL (2003). "Anticodon sequence mutants of Escherichia coli initiator tRNA: effects of overproduction of aminoacyl-tRNA synthetases, methionyl-tRNA formyltransferase, and initiation factor 2 on activity in initiation." Biochemistry 42(17);4787-99. PMID: 12718519

Mazel94: Mazel D, Pochet S, Marliere P (1994). "Genetic characterization of polypeptide deformylase, a distinctive enzyme of eubacterial translation." EMBO J 13(4);914-23. PMID: 8112305

Meinnel93: Meinnel T, Guillon JM, Mechulam Y, Blanquet S (1993). "The Escherichia coli fmt gene, encoding methionyl-tRNA(fMet) formyltransferase, escapes metabolic control." J Bacteriol 175(4);993-1000. PMID: 8432722

Meinnel93a: Meinnel T, Mechulam Y, Blanquet S (1993). "Methionine as translation start signal: a review of the enzymes of the pathway in Escherichia coli." Biochimie 75(12);1061-75. PMID: 8199241

Newton99: Newton DT, Creuzenet C, Mangroo D (1999). "Formylation is not essential for initiation of protein synthesis in all eubacteria." J Biol Chem 274(32);22143-6. PMID: 10428776

RajBhandary94: RajBhandary UL (1994). "Initiator transfer RNAs." J Bacteriol 176(3);547-52. PMID: 7507918

Ramesh97: Ramesh V, Gite S, Li Y, RajBhandary UL (1997). "Suppressor mutations in Escherichia coli methionyl-tRNA formyltransferase: role of a 16-amino acid insertion module in initiator tRNA recognition." Proc Natl Acad Sci U S A 94(25);13524-9. PMID: 9391059

Ramesh98: Ramesh V, Gite S, RajBhandary UL (1998). "Functional interaction of an arginine conserved in the sixteen amino acid insertion module of Escherichia coli methionyl-tRNA formyltransferase with determinants for formylation in the initiator tRNA." Biochemistry 37(45);15925-32. PMID: 9843398

Ramesh99: Ramesh V, Mayer C, Dyson MR, Gite S, RajBhandary UL (1999). "Induced fit of a peptide loop of methionyl-tRNA formyltransferase triggered by the initiator tRNA substrate." Proc Natl Acad Sci U S A 96(3);875-80. PMID: 9927661

Schmitt96: Schmitt E, Blanquet S, Mechulam Y (1996). "Structure of crystalline Escherichia coli methionyl-tRNA(f)Met formyltransferase: comparison with glycinamide ribonucleotide formyltransferase." EMBO J 15(17);4749-58. PMID: 8887566

Schmitt96a: Schmitt E, Mechulam Y, Ruff M, Mitschler A, Moras D, Blanquet S (1996). "Crystallization and preliminary X-ray analysis of Escherichia coli methionyl-tRNA(fMet) formyltransferase." Proteins 25(1);139-41. PMID: 8727328

Schmitt96b: Schmitt E, Guillon JM, Meinnel T, Mechulam Y, Dardel F, Blanquet S (1996). "Molecular recognition governing the initiation of translation in Escherichia coli. A review." Biochimie 78(7);543-54. PMID: 8955898

Schmitt98: Schmitt E, Panvert M, Blanquet S, Mechulam Y (1998). "Crystal structure of methionyl-tRNAfMet transformylase complexed with the initiator formyl-methionyl-tRNAfMet." EMBO J 17(23);6819-26. PMID: 9843487

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

UniProt10: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-11 released on 2010-11-02 00:00:00." Database.

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

Vaughan02: Vaughan MD, Sampson PB, Honek JF (2002). "Methionine in and out of proteins: targets for drug design." Curr Med Chem 9(3);385-409. PMID: 11860363

Wallis95: Wallis NG, Dardel F, Blanquet S (1995). "Heteronuclear NMR studies of the interactions of 15N-labeled methionine-specific transfer RNAs with methionyl-tRNA transformylase." Biochemistry 34(23);7668-77. PMID: 7779813

Zuleeg00: Zuleeg T, Vogtherr M, Schubel H, Limmer S (2000). "The C-A mismatch base pair and the single-strand terminus in the E. coli initiator tRNA(fMet) acceptor stem adopt unusual conformations." FEBS Lett 472(2-3);247-53. PMID: 10788620


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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 Nov 28, 2014, biocyc14.