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Escherichia coli K-12 substr. MG1655 Polypeptide: protein chain elongation factor EF-Ts



Gene: tsf Accession Numbers: EG11033 (EcoCyc), b0170, ECK0169

Synonyms: EF1B

Regulation Summary Diagram: ?

Summary:
EF-Ts is an elongation factor associated with the ribosome; it functions as the guanine nucleotide exchange factor for the EF-Tu GTPase. EF-Ts was first identified in the "T" (for transfer) fraction consisting of a stable (Ts) and an unstable (Tu) component [LucasLenard66]. Two molecules each of elongation factor Ts and elongation factor Tu assemble into a complex [Miller69, Wittinghofer83, Kawashima96]. EF-Ts is essential for growth [Hwang89, Baba06].

EF-Ts interacts with aminoacyl-tRNA-bound EF-Tu and regulates its affinity for GTP and aa-tRNA ligands. EF-Ts may regulate the stability of the EF-Tu-GTP-aa-tRNA ternary complex by facilitating the conformational changes within EF-Tu that are responsible for binding and release of aa-tRNAs [Burnett13].

EF-Ts enhances the rate of dissociation of GDP from EF-Tu within the EF-Tu-GDP-EF-Ts complex [Chau81, Wittinghofer83] by a factor of 6x104 [Gromadski02]. EF-Ts dissociates from the ternary EF-Tu-GDP/GTP-EF-Ts complex 103-104 times faster than from the binary EF-Tu-EF-Ts complex. Under conditions resembling those in vivo, the overall nucleotide exchange rate catalyzed by EF-Ts is 30 s-1 [Gromadski02]. The kinetics of the nucleotide exchange reaction has also been modelled [Manchester04].

A crystal structure of the EF-Tu-EF-Ts complex (EF-T) has been solved at 2.5 Å resolution. It shows two subunits each of EF-Ts and EF-Tu, where a coiled-coil motif of EF-Ts appears to be primarily responsible for dimerization [Kawashima96]. An EF-Ts mutant lacking this coiled-coil motif shows near-normal growth, but forms unstable EF-Tu-EF-Ts complexes in the presence of guanine nucleotides [Karring03].

The N-terminal part of EF-Ts is required for the nucleotide exchange activity [Bogestrand95] and appears to determine the strength of the interaction with EF-Tu [Zhang97a]. A conformational change in the N terminus may be essential for binding EF-Tu [Hwang97]. Further site-directed mutagenesis showed that residues within the N-terminal domain and subdomain C are important for interaction with EF-Tu [Zhang98b]. Residues that are thought to be involved in the guanine nucleotide exchange reaction have been mutagenized. The D80 and F81 residues are important, but not essential for activity; they appear to affect the interaction with EF-Tu [Zhang96].

The mechanism of nucleotide exchange has been investigated. Initial contact of the N-terminal domain of EF-Ts with helix D of EF-Tu appears to weaken interactions with the guanine base; contacts with the phosphate binding site, promoting release of the phosphate moiety, take place later [Wieden02].

EF-Ts enhances phosphorylation of EF-Tu [Alexander95] and can act as a "steric chaperone" for EF-Tu mutants which do not fold correctly [Krab01].

In addition to its function as a translation elongation factor, EF-Ts is a host-supplied component of the phage Qβ RNA replicase complex [Blumenthal72, Blumenthal76]. An EF-Ts mutant that forms unstable EF-Tu-EF-Ts complexes is unable to form an active Qβ-polymerase complex; EF-Ts may thus be required to stably trap EF-Tu within the Qβ-polymerase [Karring04]. Crystal structures of the Qβ replicase core complex have been solved [Kidmose10, Takeshita10].

Reviews: [LucasLenard71, Andersen03]

Citations: [Han07a, Wieden10, Pan11, Miyajima78]

Gene Citations: [An81]

Locations: cytosol, membrane

Map Position: [190,857 -> 191,708] (4.11 centisomes)
Length: 852 bp / 283 aa

Molecular Weight of Polypeptide: 30.423 kD (from nucleotide sequence)

Unification Links: ASAP:ABE-0000579 , CGSC:64 , DIP:DIP-31835N , EchoBASE:EB1026 , EcoGene:EG11033 , EcoliWiki:b0170 , Mint:MINT-1273899 , OU-Microarray:b0170 , PortEco:tsf , PR:PRO_000024132 , Pride:P0A6P1 , Protein Model Portal:P0A6P1 , RefSeq:NP_414712 , RegulonDB:EG11033 , SMR:P0A6P1 , String:511145.b0170 , UniProt:P0A6P1

Relationship Links: InterPro:IN-FAMILY:IPR000449 , InterPro:IN-FAMILY:IPR001816 , InterPro:IN-FAMILY:IPR009060 , InterPro:IN-FAMILY:IPR014039 , InterPro:IN-FAMILY:IPR018101 , Panther:IN-FAMILY:PTHR11741 , PDB:Structure:1EFU , PDB:Structure:3MMP , Pfam:IN-FAMILY:PF00627 , Pfam:IN-FAMILY:PF00889 , Prosite:IN-FAMILY:PS01126 , Prosite:IN-FAMILY:PS01127

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0043547 - positive regulation of GTPase activity Inferred from experiment [Gromadski02]
GO:0006412 - translation Inferred by computational analysis [UniProtGOA11a]
GO:0006414 - translational elongation Inferred by computational analysis [UniProtGOA11a, GOA06, GOA01a]
Molecular Function: GO:0005085 - guanyl-nucleotide exchange factor activity Inferred from experiment [Gromadski02]
GO:0005515 - protein binding Inferred from experiment [Lasserre06, Arifuzzaman06, Butland05, Kawashima96]
GO:0008270 - zinc ion binding Inferred from experiment [Katayama02]
GO:0003746 - translation elongation factor activity Inferred by computational analysis [UniProtGOA11a, GOA06, GOA01a]
Cellular Component: GO:0005737 - cytoplasm Inferred from experiment Inferred by computational analysis [UniProtGOA11, UniProtGOA11a, GOA06, Lasserre06]
GO:0005829 - cytosol Inferred from experiment Inferred by computational analysis [DiazMejia09, Ishihama08, Zhang07b, LopezCampistrou05]
GO:0016020 - membrane Inferred from experiment [Lasserre06]
GO:0005622 - intracellular Inferred by computational analysis [GOA01a]

MultiFun Terms: information transfer protein related translation

Essentiality data for tsf 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 ? 04-Apr-2013 by Keseler I , SRI International


Sequence Features

Feature Class Location Citations Comment
Cleavage-of-Initial-Methionine 1
[Pasquali94, Wilkins98, Link97, UniProt12a]
UniProt: Removed.
Chain 2 -> 283
[UniProt09]
UniProt: Elongation factor Ts;
Mutagenesis-Variant 24
[Zhang98b, UniProt11a]
Alternate sequence: K → A; UniProt: No change in binding to EF-Tu and in promoting GDP exchange.
Protein-Segment 80 -> 83
[UniProt10]
UniProt: Involved in Mg(2+) ion dislocation from EF-Tu; Sequence Annotation Type: region of interest;
Mutagenesis-Variant 81
[Zhang98b, UniProt11a]
Alternate sequence: D → A; UniProt: 2 to 3-fold less active in promoting GDP exchange and slightly lower binding constant for interaction with EF- Tu.
Mutagenesis-Variant 82
[Zhang98b, UniProt11a]
Alternate sequence: F → A; UniProt: 2 to 3-fold less active in promoting GDP exchange and 6-fold less active in binding to EF-Tu.
Acetylation-Modification 95
[Yu08a]
 
Mutagenesis-Variant 148
[Zhang98b, UniProt11a]
Alternate sequence: H → A; UniProt: At least 100-fold decrease in affinity between EF-Ts and EF-Tu and only small amount of GDP exchange activity.
Acetylation-Modification 247
[Yu08a]
 


Gene Local Context (not to scale): ?

Transcription Unit:

Notes:

History:
10/20/97 Gene b0170 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG11033; confirmed by SwissProt match.


References

Alexander95: Alexander C, Bilgin N, Lindschau C, Mesters JR, Kraal B, Hilgenfeld R, Erdmann VA, Lippmann C (1995). "Phosphorylation of elongation factor Tu prevents ternary complex formation." J Biol Chem 270(24);14541-7. PMID: 7782317

An81: An G, Bendiak DS, Mamelak LA, Friesen JD (1981). "Organization and nucleotide sequence of a new ribosomal operon in Escherichia coli containing the genes for ribosomal protein S2 and elongation factor Ts." Nucleic Acids Res 1981;9(16);4163-72. PMID: 6272196

Andersen03: Andersen GR, Nissen P, Nyborg J (2003). "Elongation factors in protein biosynthesis." Trends Biochem Sci 28(8);434-41. PMID: 12932732

Arifuzzaman06: Arifuzzaman M, Maeda M, Itoh A, Nishikata K, Takita C, Saito R, Ara T, Nakahigashi K, Huang HC, Hirai A, Tsuzuki K, Nakamura S, Altaf-Ul-Amin M, Oshima T, Baba T, Yamamoto N, Kawamura T, Ioka-Nakamichi T, Kitagawa M, Tomita M, Kanaya S, Wada C, Mori H (2006). "Large-scale identification of protein-protein interaction of Escherichia coli K-12." Genome Res 16(5);686-91. PMID: 16606699

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

Blumenthal72: Blumenthal T, Landers TA, Weber K (1972). "Bacteriophage Q replicase contains the protein biosynthesis elongation factors EF Tu and EF Ts." Proc Natl Acad Sci U S A 69(5);1313-7. PMID: 4624757

Blumenthal76: Blumenthal T, Landers TA (1976). "Renaturation of a multisubunit multiactivity enzyme complex: recovery of phage Qbeta RNA replicase, EF-Tu, and EF-Ts activities after denaturation in urea." Biochemistry 15(2);422-5. PMID: 764866

Bogestrand95: Bogestrand S, Wiborg O, Thirup S, Nyborg J (1995). "Analysis and crystallization of a 25 kDa C-terminal fragment of cloned elongation factor Ts from Escherichia coli." FEBS Lett 368(1);49-54. PMID: 7615087

Burnett13: Burnett BJ, Altman RB, Ferrao R, Alejo JL, Kaur N, Kanji J, Blanchard SC (2013). "Elongation Factor-Ts Directly Facilitates the Formation and Disassembly of the Escherichia Coli Elongation Factor-Tu-GTP-Aminoacyl-tRNA Ternary Complex." J Biol Chem. PMID: 23539628

Butland05: Butland G, Peregrin-Alvarez JM, Li J, Yang W, Yang X, Canadien V, Starostine A, Richards D, Beattie B, Krogan N, Davey M, Parkinson J, Greenblatt J, Emili A (2005). "Interaction network containing conserved and essential protein complexes in Escherichia coli." Nature 433(7025);531-7. PMID: 15690043

Chau81: Chau V, Romero G, Biltonen RL (1981). "Kinetic studies on the interactions of Escherichia coli K12 elongation factor Tu with GDP and elongation factor Ts." J Biol Chem 256(11);5591-6. PMID: 7016856

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."

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

Gromadski02: Gromadski KB, Wieden HJ, Rodnina MV (2002). "Kinetic mechanism of elongation factor Ts-catalyzed nucleotide exchange in elongation factor Tu." Biochemistry 41(1);162-9. PMID: 11772013

Han07a: Han KY, Song JA, Ahn KY, Park JS, Seo HS, Lee J (2007). "Enhanced solubility of heterologous proteins by fusion expression using stress-induced Escherichia coli protein, Tsf." FEMS Microbiol Lett 274(1);132-8. PMID: 17608803

Hwang89: Hwang YW, Sanchez A, Miller DL (1989). "Mutagenesis of bacterial elongation factor Tu at lysine 136. A conserved amino acid in GTP regulatory proteins." J Biol Chem 264(14);8304-9. PMID: 2498311

Hwang97: Hwang YW, Sanchez A, Hwang MC, Miller DL (1997). "The role of cysteinyl residues in the activity of bacterial elongation factor Ts, a guanosine nucleotide dissociation protein." Arch Biochem Biophys 348(1);157-62. PMID: 9390186

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

Karring03: Karring H, Bjornsson A, Thirup S, Clark BF, Knudsen CR (2003). "Functional effects of deleting the coiled-coil motif in Escherichia coli elongation factor Ts." Eur J Biochem 270(21);4294-305. PMID: 14622294

Karring04: Karring H, Mathu SG, van Duin J, Clark BF, Kraal B, Knudsen CR (2004). "Qbeta-phage resistance by deletion of the coiled-coil motif in elongation factor Ts." J Biol Chem 279(3);1878-84. PMID: 14583631

Katayama02: Katayama A, Tsujii A, Wada A, Nishino T, Ishihama A (2002). "Systematic search for zinc-binding proteins in Escherichia coli." Eur J Biochem 269(9);2403-13. PMID: 11985624

Kawashima96: Kawashima T, Berthet-Colominas C, Wulff M, Cusack S, Leberman R (1996). "The structure of the Escherichia coli EF-Tu.EF-Ts complex at 2.5 A resolution." Nature 379(6565);511-8. PMID: 8596629

Kidmose10: Kidmose RT, Vasiliev NN, Chetverin AB, Andersen GR, Knudsen CR (2010). "Structure of the Qbeta replicase, an RNA-dependent RNA polymerase consisting of viral and host proteins." Proc Natl Acad Sci U S A 107(24);10884-9. PMID: 20534494

Krab01: Krab IM, te Biesebeke R, Bernardi A, Parmeggiani A (2001). "Elongation factor Ts can act as a steric chaperone by increasing the solubility of nucleotide binding-impaired elongation factor-Tu." Biochemistry 40(29);8531-5. PMID: 11456491

Lasserre06: Lasserre JP, Beyne E, Pyndiah S, Lapaillerie D, Claverol S, Bonneu M (2006). "A complexomic study of Escherichia coli using two-dimensional blue native/SDS polyacrylamide gel electrophoresis." Electrophoresis 27(16);3306-21. PMID: 16858726

Link97: Link AJ, Robison K, Church GM (1997). "Comparing the predicted and observed properties of proteins encoded in the genome of Escherichia coli K-12." Electrophoresis 18(8);1259-313. PMID: 9298646

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

LucasLenard66: Lucas-Lenard J, Lipmann F (1966). "Separation of three microbial amino acid polymerization factors." Proc Natl Acad Sci U S A 55(6);1562-6. PMID: 4289971

LucasLenard71: Lucas-Lenard J (1971). "Protein biosynthesis." Annu Rev Biochem 40;409-48. PMID: 4941237

Manchester04: Manchester KL (2004). "Determination of the kinetics of guanine nucleotide exchange on EF-Tu and EF-Ts: continuing uncertainties." Biochem Biophys Res Commun 314(1);1-5. PMID: 14715237

Miller69: Miller DL, Weissbach H (1969). "An interaction between the transfer factors required for protein synthesis." Arch Biochem Biophys 132(1);146-50. PMID: 4893410

Miyajima78: Miyajima A, Kaziro Y (1978). "Coordination of levels of elongation factors Tu, Ts, and G, and ribosomal protein SI in Escherichia coli." J Biochem 83(2);453-62. PMID: 344309

Pan11: Pan JY, Wu H, Liu X, Li PP, Li H, Wang SY, Peng XX (2011). "Complexome of Escherichia coli cytosolic proteins under normal native conditions." Mol Biosyst 7(9);2651-63. PMID: 21717022

Pasquali94: Pasquali C., Sanchez J.-C., Ravier F., Golaz O., Hughes G.J., Frutiger S., Paquet N., Wilkins M., Appel R.D., Bairoch A., Hochstrasser D.F. (1994). Data submission to UniProtKB on 1994-09.

Takeshita10: Takeshita D, Tomita K (2010). "Assembly of Q{beta} viral RNA polymerase with host translational elongation factors EF-Tu and -Ts." Proc Natl Acad Sci U S A 107(36);15733-8. PMID: 20798060

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.

UniProt11a: UniProt Consortium (2011). "UniProt version 2011-06 released on 2011-06-30 00:00:00." Database.

UniProt12a: UniProt Consortium (2012). "UniProt version 2012-02 released on 2012-02-29 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."

Wieden02: Wieden HJ, Gromadski K, Rodnin D, Rodnina MV (2002). "Mechanism of elongation factor (EF)-Ts-catalyzed nucleotide exchange in EF-Tu. Contribution of contacts at the guanine base." J Biol Chem 277(8);6032-6. PMID: 11744709

Wieden10: Wieden HJ, Mercier E, Gray J, Steed B, Yawney D (2010). "A combined molecular dynamics and rapid kinetics approach to identify conserved three-dimensional communication networks in elongation factor Tu." Biophys J 99(11);3735-43. PMID: 21112298

Wilkins98: Wilkins MR, Gasteiger E, Tonella L, Ou K, Tyler M, Sanchez JC, Gooley AA, Walsh BJ, Bairoch A, Appel RD, Williams KL, Hochstrasser DF (1998). "Protein identification with N and C-terminal sequence tags in proteome projects." J Mol Biol 278(3);599-608. PMID: 9600841

Wittinghofer83: Wittinghofer A, Guariguata R, Leberman R (1983). "Bacterial elongation factor Ts: isolation and reactivity with elongation factor Tu." J Bacteriol 153(3);1266-71. PMID: 6681813

Wurmbach79: Wurmbach P, Nierhaus KH (1979). "Isolation of the protein synthesis elongation factors EF-Tu, EF-Ts, and EF-G from Escherichia coli." Methods Enzymol 60;593-606. PMID: 379535

Yu08a: Yu BJ, Kim JA, Moon JH, Ryu SE, Pan JG (2008). "The diversity of lysine-acetylated proteins in Escherichia coli." J Microbiol Biotechnol 18(9);1529-36. PMID: 18852508

Zhang07b: Zhang N, Chen R, Young N, Wishart D, Winter P, Weiner JH, Li L (2007). "Comparison of SDS- and methanol-assisted protein solubilization and digestion methods for Escherichia coli membrane proteome analysis by 2-D LC-MS/MS." Proteomics 7(4);484-93. PMID: 17309111

Zhang96: Zhang Y, Li X, Spremulli LL (1996). "Role of the conserved aspartate and phenylalanine residues in prokaryotic and mitochondrial elongation factor Ts in guanine nucleotide exchange." FEBS Lett 391(3);330-2. PMID: 8765000

Zhang97a: Zhang Y, Sun V, Spremulli LL (1997). "Role of domains in Escherichia coli and mammalian mitochondrial elongation factor Ts in the interaction with elongation factor Tu." J Biol Chem 272(35);21956-63. PMID: 9268331

Zhang98b: Zhang Y, Yu NJ, Spremulli LL (1998). "Mutational analysis of the roles of residues in Escherichia coli elongation factor Ts in the interaction with elongation factor Tu." J Biol Chem 273(8);4556-62. PMID: 9468511

Other References Related to Gene Regulation

Aseev08: Aseev LV, Levandovskaya AA, Tchufistova LS, Scaptsova NV, Boni IV (2008). "A new regulatory circuit in ribosomal protein operons: S2-mediated control of the rpsB-tsf expression in vivo." RNA 14(9):1882-94. PMID: 18648071

MendozaVargas09: Mendoza-Vargas A, Olvera L, Olvera M, Grande R, Vega-Alvarado L, Taboada B, Jimenez-Jacinto V, Salgado H, Juarez K, Contreras-Moreira B, Huerta AM, Collado-Vides J, Morett E (2009). "Genome-wide identification of transcription start sites, promoters and transcription factor binding sites in E. coli." PLoS One 4(10);e7526. PMID: 19838305


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
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