fusA

Summary

Gene Symbol: fusA
Description: protein chain elongation factor EF-G, GTP-binding
Alias: ECK3327, JW3302, far, fus
Species: Escherichia coli str. K-12 substr. MG1655

Top Publications

  1. Nakamura K, Miyamoto H, Suzuma S, Sakamoto T, Kawai G, Yamane K. Minimal functional structure of Escherichia coli 4.5 S RNA required for binding to elongation factor G. J Biol Chem. 2001;276:22844-9 pubmed
    ..These results indicate the mechanism by which EF-G interacts with 4.5 S RNA and the importance of the bulge structure for EF-G binding. ..
  2. Borowski C, Rodnina M, Wintermeyer W. Truncated elongation factor G lacking the G domain promotes translocation of the 3' end but not of the anticodon domain of peptidyl-tRNA. Proc Natl Acad Sci U S A. 1996;93:4202-6 pubmed
    ..The G domain's intramolecular interactions are likely to be modulated by GTP binding and hydrolysis. ..
  3. Agrawal R, Sharma M, Kiel M, Hirokawa G, Booth T, Spahn C, et al. Visualization of ribosome-recycling factor on the Escherichia coli 70S ribosome: functional implications. Proc Natl Acad Sci U S A. 2004;101:8900-5 pubmed publisher
  4. Agrawal R, Heagle A, Penczek P, Grassucci R, Frank J. EF-G-dependent GTP hydrolysis induces translocation accompanied by large conformational changes in the 70S ribosome. Nat Struct Biol. 1999;6:643-7 pubmed
    ..Upon GTP hydrolysis, the bifurcation is reversed and an arc-like connection is formed between the base of the stalk and EF-G. ..
  5. Karimi R, Pavlov M, Buckingham R, Ehrenberg M. Novel roles for classical factors at the interface between translation termination and initiation. Mol Cell. 1999;3:601-9 pubmed
    ..We show that this step requires initiation factor IF3, whose role was previously thought to be restricted to promoting specific 30S initiation complex formation from free 30S subunits. ..
  6. Seo H, Kiel M, Pan D, Raj V, Kaji A, Cooperman B. Kinetics and thermodynamics of RRF, EF-G, and thiostrepton interaction on the Escherichia coli ribosome. Biochemistry. 2004;43:12728-40 pubmed
    ..Added thiostrepton slows the initial binding of EF-G, and prevents both formation of the more stable EF-G complex and EF-G-induced RRF dissociation. These findings are relevant for the mechanism of post-termination complex disassembly. ..
  7. Marshall R, Dorywalska M, Puglisi J. Irreversible chemical steps control intersubunit dynamics during translation. Proc Natl Acad Sci U S A. 2008;105:15364-9 pubmed publisher
    ..Intersubunit rotation is likely a requirement for the concerted movement of tRNA and mRNA substrates during translocation. ..
  8. Spiegel P, Ermolenko D, Noller H. Elongation factor G stabilizes the hybrid-state conformation of the 70S ribosome. RNA. 2007;13:1473-82 pubmed
    ..Our results show that binding of EF-G in the presence of GDPNP or GDP.fusidic acid stabilizes the ribosomal intermediate hybrid state, but that complete translocation is supported only by EF-G.GTP or EF-G.GDPNP. ..
  9. Ito K, Fujiwara T, Toyoda T, Nakamura Y. Elongation factor G participates in ribosome disassembly by interacting with ribosome recycling factor at their tRNA-mimicry domains. Mol Cell. 2002;9:1263-72 pubmed
    ..These mutational studies suggest that EF-G motor action is transmitted to RRF by specific surface contacts between the domains that mimic the anticodon arm...

More Information

Publications70

  1. Peske F, Savelsbergh A, Katunin V, Rodnina M, Wintermeyer W. Conformational changes of the small ribosomal subunit during elongation factor G-dependent tRNA-mRNA translocation. J Mol Biol. 2004;343:1183-94 pubmed
  2. Bowen W, Van Dyke N, Murgola E, Lodmell J, Hill W. Interaction of thiostrepton and elongation factor-G with the ribosomal protein L11-binding domain. J Biol Chem. 2005;280:2934-43 pubmed
    ..The implications for recent models of thiostrepton activity and factor interactions are discussed. ..
  3. Alakhov Y, Dovgas N, Motuz L, Vinokurov L, Ovchinnikov Y. The primary structure of the elongation factor G from Escherichia coli: amino acid sequence of the C-terminal domain. FEBS Lett. 1981;126:183-6 pubmed
  4. Ganoza M, Aoki H, Burkhardt N, Murphy B. The ribosome as affinity matrix': efficient purification scheme for translation factors. Biochimie. 1996;78:51-61 pubmed
    ..The procedure markedly simplifies the isolation, in homogeneous form, of all the non-ribosomal proteins required to reconstruct translation. ..
  5. Saito K, Mattheakis L, Nomura M. Post-transcriptional regulation of the str operon in Escherichia coli. Ribosomal protein S7 inhibits coupled translation of S7 but not its independent translation. J Mol Biol. 1994;235:111-24 pubmed
    ..of Escherichia coli consists of the genes for ribosomal proteins S12 (rpsL) and S7 (rpsG) and elongation factors G (fusA) and Tu (tufA)...
  6. Nakamura K, Fujii Y, Shibata T, Yamane K. Depletion of Escherichia coli 4.5S RNA leads to an increase in the amount of protein elongation factor EF-G associated with ribosomes. Eur J Biochem. 1999;259:543-50 pubmed
    ..5S RNA is concerned with the mode of association of EF-G with the ribosomes. Moreover, this function depends on the secondary structure of 4.5S RNA as well as a ten-base sequence conserved between the two RNAs. ..
  7. Fujiwara T, Ito K, Yamami T, Nakamura Y. Ribosome recycling factor disassembles the post-termination ribosomal complex independent of the ribosomal translocase activity of elongation factor G. Mol Microbiol. 2004;53:517-28 pubmed
    ..These findings suggest that RRF is not a functional mimic of tRNA and disassembles the post-termination ribosomal complex independently of the translocation activity of EF-G. ..
  8. Kaji A, Kiel M, Hirokawa G, Muto A, Inokuchi Y, Kaji H. The fourth step of protein synthesis: disassembly of the posttermination complex is catalyzed by elongation factor G and ribosome recycling factor, a near-perfect mimic of tRNA. Cold Spring Harb Symp Quant Biol. 2001;66:515-29 pubmed
  9. Nechifor R, Murataliev M, Wilson K. Functional interactions between the G' subdomain of bacterial translation factor EF-G and ribosomal protein L7/L12. J Biol Chem. 2007;282:36998-7005 pubmed
    ..Together, these results provide evidence for functionally important interactions between helix AG' of EF-G and L7/L12 of the ribosome. ..
  10. Konevega A, Fischer N, Semenkov Y, Stark H, Wintermeyer W, Rodnina M. Spontaneous reverse movement of mRNA-bound tRNA through the ribosome. Nat Struct Mol Biol. 2007;14:318-24 pubmed
    ..These results lend support to the diffusion model of tRNA movement during translocation. In the cell, tRNA movement is biased in the forward direction by EF-G, which acts as a Brownian ratchet and prevents backward movement. ..
  11. Kraal B, Bosch L, Mesters J, de Graaf J, Woudt L, Vijgenboom E, et al. Elongation factors in protein synthesis. Ciba Found Symp. 1993;176:28-43; discussion 44-52 pubmed
    ..We have found a synergistic effect between EF-Tu and EF-G in their uncoupled GTPase reactions on empty ribosomes. Only the EF-G reaction is perturbed by fluoroaluminates. ..
  12. Sergiev P, Lesnyak D, Kiparisov S, Burakovsky D, Leonov A, Bogdanov A, et al. Function of the ribosomal E-site: a mutagenesis study. Nucleic Acids Res. 2005;33:6048-56 pubmed
    ..The mutation affects formation of the P/E hybrid site and leads to a loss of stimulation of the multiple turnover GTPase activity of EF-G by deacylated tRNA bound to the ribosome. ..
  13. Ticu C, Murataliev M, Nechifor R, Wilson K. A central interdomain protein joint in elongation factor G regulates antibiotic sensitivity, GTP hydrolysis, and ribosome translocation. J Biol Chem. 2011;286:21697-705 pubmed publisher
    ..These experimental results reveal the multipurpose roles of an interdomain joint in the heart of an essential translation factor that can both promote and inhibit bacterial translation. ..
  14. Caldon C, Yoong P, March P. Evolution of a molecular switch: universal bacterial GTPases regulate ribosome function. Mol Microbiol. 2001;41:289-97 pubmed
    ..If this is the case, then a corollary of this hypothesis is that GTPases that do not bind RNA arose at a later time from an RNA-binding progenitor that lost the capability to bind RNA. ..
  15. Hou Y, Lin Y, Sharer J, March P. In vivo selection of conditional-lethal mutations in the gene encoding elongation factor G of Escherichia coli. J Bacteriol. 1994;176:123-9 pubmed
    ..this process, a genetic selection was developed with the aim of obtaining conditional-lethal alleles of the gene (fusA) that encodes EF-G in Escherichia coli...
  16. Sharma H, Adio S, Senyushkina T, Belardinelli R, Peske F, Rodnina M. Kinetics of Spontaneous and EF-G-Accelerated Rotation of Ribosomal Subunits. Cell Rep. 2016;16:2187-2196 pubmed publisher
    ..The transition back to the non-rotated state of the ribosome kinetically coincides with tRNA-mRNA movement. Thus, large-scale movements of the ribosome are intrinsically rapid and gated by its ligands such as EF-G and tRNA. ..
  17. Rezuchova B, Miticka H, Homerova D, Roberts M, Kormanec J. New members of the Escherichia coli sigmaE regulon identified by a two-plasmid system. FEMS Microbiol Lett. 2003;225:1-7 pubmed
    ..The new rpoE-dependent promoters governed expression of genes encoding proteins involved in primary metabolism (fusA, tufA, recR), phospholipid and lipopolysaccharide biosynthesis (psd, lpxP), signal transduction (sixA), proposed ..
  18. Baxter R, Ganoza M, Zahid N, Chung D. Reconstruction of peptidyltransferase activity on 50S and 70S ribosomal particles by peptide fragments of protein L16. Eur J Biochem. 1987;163:473-9 pubmed
    ..Also, EF-P, a soluble protein which reconstructs the peptide-bond formation and transesterification reactions on 70S ribosomes, stimulated both peptidyltransferase activities exhibited by the L16 N-terminal peptide. ..
  19. Caldas T, Laalami S, Richarme G. Chaperone properties of bacterial elongation factor EF-G and initiation factor IF2. J Biol Chem. 2000;275:855-60 pubmed
    ..These results suggest that EF-G and IF2, in addition to their role in translation, might be implicated in protein folding and protection from stress. ..
  20. Wang Y, Qin H, Kudaravalli R, Kirillov S, Dempsey G, Pan D, et al. Single-molecule structural dynamics of EF-G--ribosome interaction during translocation. Biochemistry. 2007;46:10767-75 pubmed
    ..6 state results from the conformational lability of unlocked ribosomes formed during translocation. An idling state, possibly pertinent to regulation of protein synthesis, is detected in some ribosomes in the poly(Phe) system. ..
  21. Pavlov M, Freistroffer D, Heurgué Hamard V, Buckingham R, Ehrenberg M. Release factor RF3 abolishes competition between release factor RF1 and ribosome recycling factor (RRF) for a ribosome binding site. J Mol Biol. 1997;273:389-401 pubmed
    ..The overproduction of RF1 in cells deficient in RRF or lacking RF3 has effects on growth rate predicted by the in vitro experiments. ..
  22. Gao H, Valle M, Ehrenberg M, Frank J. Dynamics of EF-G interaction with the ribosome explored by classification of a heterogeneous cryo-EM dataset. J Struct Biol. 2004;147:283-90 pubmed
    ..The possible presence of additional intermediate states is discussed. ..
  23. Hamel E, Koka M, Nakamoto T. Requirement of an Escherichia coli 50 S ribosomal protein component for effective interaction of the ribosome with T and G factors and with guanosine triphosphate. J Biol Chem. 1972;247:805-14 pubmed
  24. Voigt J, Nagel K. Isolation and characterization of an inhibitor of ribosome-dependent GTP hydrolysis by elongation factor G. Eur J Biochem. 1990;194:579-85 pubmed
    ..When ribosomes were preincubated with poly(U) and Phe-tRNA(Phe), poly(phenylalanine) synthesis was considerably less affected by the inhibitor, whereas EF-G GTPase was still sensitive. ..
  25. Yokota T, Sugisaki H, Takanami M, Kaziro Y. The nucleotide sequence of the cloned tufA gene of Escherichia coli. Gene. 1980;12:25-31 pubmed
    ..The nucleotide sequence of about 1500 base pairs, covering the C-terminal portion of elongation factor EF-G (fus gene), the intercistronic region between fus and tufA, the entire structural gene for tufA with the GUG initiation ..
  26. Kischa K, Moller W, Stoffler G. Reconstitution of a GTPase activity by a 50S ribosomal protein and E. coli. Nat New Biol. 1971;233:62-3 pubmed
  27. Qin B, Yamamoto H, Ueda T, Varshney U, Nierhaus K. The Termination Phase in Protein Synthesis is not Obligatorily Followed by the RRF/EF-G-Dependent Recycling Phase. J Mol Biol. 2016;428:3577-87 pubmed publisher
    ..In this view, recycling after termination is a special case of the general RRF function, which happens whenever fMet-tRNA is not available for triggering 70S scanning. ..
  28. Sharer J, Koosha H, Church W, March P. The function of conserved amino acid residues adjacent to the effector domain in elongation factor G. Proteins. 1999;37:293-302 pubmed
    ..acid positions necessary for EF-G functions, a series of mutations were constructed in the EF-G structural gene (fusA) of Escherichia coli, specifically at positions flanking the effector domain...
  29. Savelsbergh A, Mohr D, Kothe U, Wintermeyer W, Rodnina M. Control of phosphate release from elongation factor G by ribosomal protein L7/12. EMBO J. 2005;24:4316-23 pubmed
  30. Chan Y, Correll C, Wool I. The location and the significance of a cross-link between the sarcin/ricin domain of ribosomal RNA and the elongation factor-G. J Mol Biol. 2004;337:263-72 pubmed
    ..The proximity suggests that the SRD RNA has a role in the activation of GTP hydrolysis that leads to a transition in the conformation of the factor and to its release from the ribosome. ..
  31. Ramrath D, Yamamoto H, Rother K, Wittek D, Pech M, Mielke T, et al. The complex of tmRNA-SmpB and EF-G on translocating ribosomes. Nature. 2012;485:526-9 pubmed publisher
  32. Johanson U, Hughes D. Comparison of the complete sequence of the str operon in Salmonella typhimurium and Escherichia coli. Gene. 1992;120:93-8 pubmed
    ..A trend towards a higher G+C content in fusA (gene encoding elongation factor (EF)-G) and tufA (gene encoding EF-Tu) in S. typhimurium is noted...
  33. Wintermeyer W, Savelsbergh A, Semenkov Y, Katunin V, Rodnina M. Mechanism of elongation factor G function in tRNA translocation on the ribosome. Cold Spring Harb Symp Quant Biol. 2001;66:449-58 pubmed
  34. Kjeldgaard M, Nyborg J, Clark B. The GTP binding motif: variations on a theme. FASEB J. 1996;10:1347-68 pubmed
    ..Finally, we discuss the important structures of complexes between GTP binding proteins and their substrates that have appeared in the literature recently. ..
  35. Nyborg J, Kjeldgaard M. Elongation in bacterial protein biosynthesis. Curr Opin Biotechnol. 1996;7:369-75 pubmed
    ..GTP and of the complex EF-Tu.EF-Ts have been determined. Within the same period, new cryo-electron microscopy reconstructions of ribosome particles have been obtained. ..
  36. Savelsbergh A, Matassova N, Rodnina M, Wintermeyer W. Role of domains 4 and 5 in elongation factor G functions on the ribosome. J Mol Biol. 2000;300:951-61 pubmed
    ..These mutants are not released from the ribosome after GTP hydrolysis or translocation, indicating that the contact with, or a conformational change of, the alpha-sarcin stem-loop is required for EF-G release from the ribosome. ..
  37. Kuwano M, Schlessinger D. G factor mutants of Escherichia coli: map location and properties. Biochem Biophys Res Commun. 1971;42:441-4 pubmed
  38. Diaconu M, Kothe U, Schl nzen F, Fischer N, Harms J, Tonevitsky A, et al. Structural basis for the function of the ribosomal L7/12 stalk in factor binding and GTPase activation. Cell. 2005;121:991-1004 pubmed publisher
    ..Highly mobile L7/12 C-terminal domains promote recruitment of translation factors to the ribosome and stimulate GTP hydrolysis by the ribosome bound factors through stabilization of their active GTPase conformation...
  39. Highland J, Ochsner E, Gordon J, Bodley J, Hasenbank R, Stoffler G. Coordinate inhibition of elongation factor G function and ribosomal subunit association by antibodies to several ribosomal proteins. Proc Natl Acad Sci U S A. 1974;71:627-30 pubmed
  40. Post L, Arfsten A, Reusser F, Nomura M. DNA sequences of promoter regions for the str and spc ribosomal protein operons in E. coli. Cell. 1978;15:215-29 pubmed
    ..Extensive sequence similarity between the str and spc promoter regions is found downstream from the Pribnow box-that is, in a transcribed region preceding the translation start sites. ..
  41. Robertson E, Aggison L, Nicholson A. Phosphorylation of elongation factor G and ribosomal protein S6 in bacteriophage T7-infected Escherichia coli. Mol Microbiol. 1994;11:1045-57 pubmed
    ..Finally, several experimental variables are analysed which influence the production and pattern of phosphorylated proteins in both uninfected and T7-infected cells. ..
  42. Ticu C, Nechifor R, Nguyen B, Desrosiers M, Wilson K. Conformational changes in switch I of EF-G drive its directional cycling on and off the ribosome. EMBO J. 2009;28:2053-65 pubmed publisher
    ..Collectively, our results support a central role of sw1 in driving the EF-G cycle during protein synthesis. ..
  43. Ishino T, Atarashi K, Uchiyama S, Yamami T, Saihara Y, Yoshida T, et al. Interaction of ribosome recycling factor and elongation factor EF-G with E. coli ribosomes studied by the surface plasmon resonance technique. Genes Cells. 2000;5:953-63 pubmed
    ..The BIACORE technique proved useful for real-time monitoring of the interaction between the ribosome and translation factors, as well as for screening of potential inhibitors for ribosome recycling factor. ..
  44. Mohr D, Wintermeyer W, Rodnina M. GTPase activation of elongation factors Tu and G on the ribosome. Biochemistry. 2002;41:12520-8 pubmed
    ..This effect appears to be augmented by interactions of other structural elements of the large ribosomal subunit with the switch regions of the factors. ..
  45. Caldas T, El Yaagoubi A, Kohiyama M, Richarme G. Purification of elongation factors EF-Tu and EF-G from Escherichia coli by covalent chromatography on thiol-sepharose. Protein Expr Purif. 1998;14:65-70 pubmed
    ..The specific reactivities of the elongation factors with thiol-Sepharose allow their efficient purification and suggest that they possess hitherto undiscovered properties connected with their reactive thiols. ..
  46. Chen J, Petrov A, Tsai A, O Leary S, Puglisi J. Coordinated conformational and compositional dynamics drive ribosome translocation. Nat Struct Mol Biol. 2013;20:718-27 pubmed publisher
    During translation elongation, the ribosome compositional factors elongation factor G (EF-G; encoded by fusA) and tRNA alternately bind to the ribosome to direct protein synthesis and regulate the conformation of the ribosome...
  47. Sagar M, Lucast L, Doudna J. Conserved but nonessential interaction of SRP RNA with translation factor EF-G. RNA. 2004;10:772-8 pubmed
    ..5S RNA, implying that cells can survive 4.5S RNA depletion by decreasing the affinity between 4.5S RNA and the translational machinery. These data suggest that SRP function is the essential role of 4.5S RNA in bacteria. ..
  48. Post L, Nomura M. DNA sequences from the str operon of Escherichia coli. J Biol Chem. 1980;255:4660-6 pubmed
    ..contains genes for ribosomal proteins (r-proteins) S12 (str or rpsL) and S7 (rpsG) and elongation factors G (fus) and Tu (tufA)...
  49. Miyajima A, Kaziro Y. Coordination of levels of elongation factors Tu, Ts, and G, and ribosomal protein SI in Escherichia coli. J Biochem. 1978;83:453-62 pubmed
    ..These results indicate that in E. coli the syntheses of the above four proteins are regulated in a coordinated manner and are subject to stringent control. ..
  50. Ganoza M, Cunningham C, Green R. A new factor from Escherichia coli affects translocation of mRNA. J Biol Chem. 1995;270:26377-81 pubmed
    ..We propose that W functions by ejecting tRNAs from ribosomes in a step that precedes the movement of mRNA during translocation. ..
  51. Highland J, Bodley J, Gordon J, Hasenbank R, Stoffler G. Identity of the ribosomal proteins involved in the interaction with elongation factor G. Proc Natl Acad Sci U S A. 1973;70:147-50 pubmed
    ..The present data are consistent with the view that the interaction between ribosomes and EF-G that results in GTP hydrolysis occurs on, and is limited to, proteins L7 and L12 on the surface of the 50S ribosomal subunit. ..
  52. Furano A, Sullivan M. The peptide chain elongation factor genes tufA and fus of Escherichia coli are intimately related physically. J Biol Chem. 1978;253:8351-4 pubmed
    ..molar concentration of this factor is considerably higher than that of elongation factor G which is encoded by the fus gene...
  53. Ishibashi M, Nakamura K, Yamane K. The functional analysis of 4.5S RNA in ribosomal translocation. Nucleic Acids Symp Ser. 1999;:199-200 pubmed
    ..5S RNA have the ability of binding EF-G using gel mobility shift assay. Increasing in the concentration of GDP increase the binding activity of 4.5S RNA to EF-G. Based on these data, we propose that 4.5S RNA release EF-G from ribosome. ..
  54. Savelsbergh A, Mohr D, Wilden B, Wintermeyer W, Rodnina M. Stimulation of the GTPase activity of translation elongation factor G by ribosomal protein L7/12. J Biol Chem. 2000;275:890-4 pubmed
    ..L7/12 appears to function by stabilizing the GTPase transition state of EF-G. ..
  55. Ovchinnikov YuA -, Alakhov YuB -, Bundulis YuP -, Bundule M, Dovgas N, Kozlov V, et al. The primary structure of elongation factor G from Escherichia coli. A complete amino acid sequence. FEBS Lett. 1982;139:130-5 pubmed
  56. Clark B, Thirup S, Kjeldgaard M, Nyborg J. Structural information for explaining the molecular mechanism of protein biosynthesis. FEBS Lett. 1999;452:41-6 pubmed
    ..Other translation factors such as the initiation factor-2 and the release factor 1 or 2 are also thought to mimic tRNA. These observations raise questions concerning the possible evolution of G-proteins involved in protein biosynthesis. ..
  57. Gottesman S. Protecting the neighborhood: extreme measures. Proc Natl Acad Sci U S A. 1998;95:2731-2 pubmed
  58. Zengel J, Archer R, Lindahl L. The nucleotide sequence of the Escherichia coli fus gene, coding for elongation factor G. Nucleic Acids Res. 1984;12:2181-92 pubmed
    We have determined the nucleotide sequence of the Escherichia coli fus gene, which codes for elongation factor G. The protein product of the sequenced gene contains 703 amino acids, with a predicted molecular weight of 77,444...
  59. Studer S, Feinberg J, Joseph S. Rapid kinetic analysis of EF-G-dependent mRNA translocation in the ribosome. J Mol Biol. 2003;327:369-81 pubmed
  60. Nechifor R, Wilson K. Crosslinking of translation factor EF-G to proteins of the bacterial ribosome before and after translocation. J Mol Biol. 2007;368:1412-25 pubmed
    ..This mechanism may ensure that a single GTP molecule is hydrolyzed for each tRNA/mRNA translocation event. ..
  61. Yu H, Chan Y, Wool I. The identification of the determinants of the cyclic, sequential binding of elongation factors tu and g to the ribosome. J Mol Biol. 2009;386:802-13 pubmed publisher