Genomes and Genes
peptide elongation factor g
Summary: Peptide Elongation Factor G catalyzes the translocation of peptidyl-tRNA from the A to the P site of bacterial ribosomes by a process linked to hydrolysis of GTP to GDP.
- Frank J, Gao H, Sengupta J, Gao N, Taylor D. The process of mRNA-tRNA translocation. Proc Natl Acad Sci U S A. 2007;104:19671-8 pubmed..New results obtained by cryoelectron microscopy, interpreted in the light of x-ray structures and kinetic data, allow us to develop a model of the molecular events during translocation. ..
- Liu H, Chen C, Zhang H, Kaur J, Goldman Y, Cooperman B. The conserved protein EF4 (LepA) modulates the elongation cycle of protein synthesis. Proc Natl Acad Sci U S A. 2011;108:16223-8 pubmed publisher..Our present results strongly suggest that PRE complex is the principal target of EF4 action on translation, rather than POST complex as had been previously supposed. ..
- Cox G, Thompson G, Jenkins H, Peske F, Savelsbergh A, Rodnina M, et al. Ribosome clearance by FusB-type proteins mediates resistance to the antibiotic fusidic acid. Proc Natl Acad Sci U S A. 2012;109:2102-7 pubmed publisher..Ribosome clearance by these proteins represents a highly unusual antibiotic resistance mechanism, which appears to be fine-tuned by the relative abundance of FusB-type protein, ribosomes, and EF-G. ..
- Tama F, Miyashita O, Brooks C. Normal mode based flexible fitting of high-resolution structure into low-resolution experimental data from cryo-EM. J Struct Biol. 2004;147:315-26 pubmed
- Zavialov A, Ehrenberg M. Peptidyl-tRNA regulates the GTPase activity of translation factors. Cell. 2003;114:113-22 pubmed..We also propose a model for translocation of tRNAs in two separate steps, which clarifies the roles of EF-G.GTP and GTP hydrolysis in this process. ..
- Shoji S, Walker S, Fredrick K. Ribosomal translocation: one step closer to the molecular mechanism. ACS Chem Biol. 2009;4:93-107 pubmed publisher..We highlight recent progress toward elucidating the molecular basis of translocation and how various antibiotics influence tRNA-mRNA movement. ..
- Peske F, Rodnina M, Wintermeyer W. Sequence of steps in ribosome recycling as defined by kinetic analysis. Mol Cell. 2005;18:403-12 pubmed..IF3 is required for the subsequent ejection of tRNA and mRNA from the small subunit. The latter step is slower than subunit dissociation and constitutes the rate-limiting step of ribosome recycling. ..
- Southworth D, Brunelle J, Green R. EFG-independent translocation of the mRNA:tRNA complex is promoted by modification of the ribosome with thiol-specific reagents. J Mol Biol. 2002;324:611-23 pubmed..These data suggest that molecular targets (ribosomal proteins) in the interface region of the ribosome are critical barriers that influence the translocation of the mRNA:tRNA complex. ..
- Yamami T, Ito K, Fujiwara T, Nakamura Y. Heterologous expression of Aquifex aeolicus ribosome recycling factor in Escherichia coli is dominant lethal by forming a complex that lacks functional co-ordination for ribosome disassembly. Mol Microbiol. 2005;55:150-61 pubmed..coli. These aaRRF mutations are spatially distinct from mutations previously described and suggest a novel active centre for coupling EF-G's G domain motor action to ribosome disassembly. ..
- Wilson D, Blaha G, Connell S, Ivanov P, Jenke H, Stelzl U, et al. Protein synthesis at atomic resolution: mechanistics of translation in the light of highly resolved structures for the ribosome. Curr Protein Pept Sci. 2002;3:1-53 pubmed..Here we try a systematic synopsis of these ribosomal functions in light of the cryo-electron microscopic structures (resolution >7 A) and the atomic x-ray structures (>2.4 A) of the ribosome. ..
- Rodnina M, Savelsbergh A, Matassova N, Katunin V, Semenkov Y, Wintermeyer W. Thiostrepton inhibits the turnover but not the GTPase of elongation factor G on the ribosome. Proc Natl Acad Sci U S A. 1999;96:9586-90 pubmed..The results indicate that thiostrepton inhibits a structural transition of the 1067 region of 23S rRNA that is important for functions of EF-G after GTP hydrolysis. ..
- Orzechowski M, Tama F. Flexible fitting of high-resolution x-ray structures into cryoelectron microscopy maps using biased molecular dynamics simulations. Biophys J. 2008;95:5692-705 pubmed publisher..In addition, we show that overfitting can be avoided by assessing the quality of the fitted model in terms of correlation coefficient and secondary structure preservation. ..
- Besier S, Ludwig A, Brade V, Wichelhaus T. Molecular analysis of fusidic acid resistance in Staphylococcus aureus. Mol Microbiol. 2003;47:463-9 pubmed..Thus, the data presented provide evidence for the crucial importance of individual amino acid exchanges within EF-G for the generation of fusidic acid resistance in S. aureus. ..
- Bhargava K, Templeton P, Spremulli L. Expression and characterization of isoform 1 of human mitochondrial elongation factor G. Protein Expr Purif. 2004;37:368-76 pubmed..Human EF-G1(mt) is considerably more resistant to fusidic acid than many bacterial translocases. A molecular model for EF-G1(mt) has been created and analyzed in the context of its relationship to the translocases from other systems. ..
- Turnidge J, Collignon P. Resistance to fusidic acid. Int J Antimicrob Agents. 1999;12 Suppl 2:S35-44 pubmed..However, evidence suggests that it does not occur at high frequency in clinical practice. Nevertheless, accumulated experience is that fusidic acid in combination with other agents results in less resistance emergence. ..
- Chen Y, Koripella R, Sanyal S, Selmer M. Staphylococcus aureus elongation factor G--structure and analysis of a target for fusidic acid. FEBS J. 2010;277:3789-803 pubmed publisher..thermophilus EF-G in complex with the 70S ribosome with fusidic acid [Gao YG et al. (2009) Science326, 694-699]. The mutations can be classified as affecting FA binding, EF-G-ribosome interactions, EF-G conformation, and EF-G stability. ..
- Cukras A, Southworth D, Brunelle J, Culver G, Green R. Ribosomal proteins S12 and S13 function as control elements for translocation of the mRNA:tRNA complex. Mol Cell. 2003;12:321-8 pubmed..These data support a model where S12 and S13 function as control elements for the more ancient rRNA- and tRNA-driven movements of translocation. ..
- Chen H, Hung W, Tseng S, Tsai J, Hsueh P, Teng L. Fusidic acid resistance determinants in Staphylococcus aureus clinical isolates. Antimicrob Agents Chemother. 2010;54:4985-91 pubmed publisher..Taken together, the distribution of fusidic acid resistance determinants (fusA mutations, fusB, and fusC) was quite different between MRSA and MSSA groups. ..
- Aevarsson A, Brazhnikov E, Garber M, Zheltonosova J, Chirgadze Y, Al Karadaghi S, et al. Three-dimensional structure of the ribosomal translocase: elongation factor G from Thermus thermophilus. EMBO J. 1994;13:3669-77 pubmed..Domains III and V show structural similarities to ribosomal proteins. Domain IV protrudes from the main body of the protein and has an extraordinary topology with a left-handed cross-over connection between two parallel beta-strands...
- Wilson K, Nechifor R. Interactions of translational factor EF-G with the bacterial ribosome before and after mRNA translocation. J Mol Biol. 2004;337:15-30 pubmed..The effects in H95 suggest that EF-G interacts weakly with H95 before mRNA translocation and strongly and more extensively with this helix following mRNA translocation. ..
- Selmer M, Al Karadaghi S, Hirokawa G, Kaji A, Liljas A. Crystal structure of Thermotoga maritima ribosome recycling factor: a tRNA mimic. Science. 1999;286:2349-52 pubmed..The structural arrangement of this mimicry is entirely different from that of other cases of less pronounced mimicry of tRNA so far described...
- 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. ..
- Avarsson A. Structure-based sequence alignment of elongation factors Tu and G with related GTPases involved in translation. J Mol Evol. 1995;41:1096-104 pubmed
- 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. ..
- Mikolajka A, Liu H, Chen Y, Starosta A, Marquez V, Ivanova M, et al. Differential effects of thiopeptide and orthosomycin antibiotics on translational GTPases. Chem Biol. 2011;18:589-600 pubmed publisher..Collectively, these results shed insight not only into fundamental aspects of translation but also into the unappreciated specificities of these classes of translational inhibitors. ..
- Antonicka H, Sasarman F, Kennaway N, Shoubridge E. The molecular basis for tissue specificity of the oxidative phosphorylation deficiencies in patients with mutations in the mitochondrial translation factor EFG1. Hum Mol Genet. 2006;15:1835-46 pubmed..Our results demonstrate marked differences among tissues in the organization of the mitochondrial translation system and its response to dysfunction, and explain the severe hepatopathy, but normal cardiac function in EFG1 patients. ..
- Voigt J, Nagel K. Regulation of elongation factor G GTPase activity by the ribosomal state. The effects of initiation factors and differentially bound tRNA, aminoacyl-tRNA, and peptidyl-tRNA. J Biol Chem. 1993;268:100-6 pubmed..The same effects are observed for Met-tRNA, Phe-tRNAPhe and uncharged tRNA. These findings are discussed in the light of the three-site model of the ribosome and the mechanism of translocation. ..
- Hanson C, Fucini P, Ilag L, Nierhaus K, Robinson C. Dissociation of intact Escherichia coli ribosomes in a mass spectrometer. Evidence for conformational change in a ribosome elongation factor G complex. J Biol Chem. 2003;278:1259-67 pubmed..These results allow us to propose that the ribosome elongation factor-G complex inhibited by thiostrepton, but not fusidic acid, involves destabilization of 5 S RNA-protein interactions. ..
- 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...
- 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. ..
- Bielecki P, Lukat P, Hüsecken K, Dötsch A, Steinmetz H, Hartmann R, et al. Mutation in elongation factor G confers resistance to the antibiotic argyrin in the opportunistic pathogen Pseudomonas aeruginosa. Chembiochem. 2012;13:2339-45 pubmed publisher..aeruginosa, analysis of the sequence identity in EF-G and its correlation with argyrin resistance in different bacteria imply that additional factors such as uptake of argyrin play a role in the argyrin resistance of other organisms. ..
- 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. ..
- Connell S, Takemoto C, Wilson D, Wang H, Murayama K, Terada T, et al. Structural basis for interaction of the ribosome with the switch regions of GTP-bound elongation factors. Mol Cell. 2007;25:751-64 pubmed..Therefore, a network of interaction with the ribosome establishes the active GTP conformation of EF-G and thus facilitates GTP hydrolysis and tRNA translocation. ..
- Burdett V. Tet(M)-promoted release of tetracycline from ribosomes is GTP dependent. J Bacteriol. 1996;178:3246-51 pubmed..Furthermore, while Tet(M) protects translation from tetracycline inhibition in a defined system, it is unable to substitute for either EF-G or elongation factor Tu. ..
- Horan L, Noller H. Intersubunit movement is required for ribosomal translocation. Proc Natl Acad Sci U S A. 2007;104:4881-5 pubmed..These findings show that intersubunit movement is required for ribosomal translocation, accounting for the universal two-subunit architecture of ribosomes. ..
- Kiel M, Raj V, Kaji H, Kaji A. Release of ribosome-bound ribosome recycling factor by elongation factor G. J Biol Chem. 2003;278:48041-50 pubmed..Before release from the ribosome, the position of RRF on the ribosome will change from the original A/P site to a new location that clashes with tRNA on the P/E site. ..
- Qin Y, Polacek N, Vesper O, Staub E, Einfeldt E, Wilson D, et al. The highly conserved LepA is a ribosomal elongation factor that back-translocates the ribosome. Cell. 2006;127:721-33 pubmed..We suggest renaming LepA as elongation factor 4 (EF4). ..
- 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. ..
- Suematsu T, Yokobori S, Morita H, Yoshinari S, Ueda T, Kita K, et al. A bacterial elongation factor G homologue exclusively functions in ribosome recycling in the spirochaete Borrelia burgdorferi. Mol Microbiol. 2010;75:1445-54 pubmed publisher..These results indicate that two B. burgdorferi EF-G paralogues are close relatives to mitochondrial EF-G paralogues rather than the conventional bacterial EF-G, in both their phylogenetic and biochemical features...
- Wriggers W, Agrawal R, Drew D, McCammon A, Frank J. Domain motions of EF-G bound to the 70S ribosome: insights from a hand-shaking between multi-resolution structures. Biophys J. 2000;79:1670-8 pubmed..The rearrangement of EF-G's structurally preserved regions, mediated and guided by flexible linkers, defines the site of interaction with the GTPase-associated center of the ribosome. ..
- Hauryliuk V, Mitkevich V, Eliseeva N, Petrushanko I, Ehrenberg M, Makarov A. The pretranslocation ribosome is targeted by GTP-bound EF-G in partially activated form. Proc Natl Acad Sci U S A. 2008;105:15678-83 pubmed publisher..We propose that the active overall conformation of EF-G is attained only in complex with the ribosome in its "ratcheted state," with hybrid tRNA binding sites. ..
- Gao Y, Selmer M, Dunham C, Weixlbaumer A, Kelley A, Ramakrishnan V. The structure of the ribosome with elongation factor G trapped in the posttranslocational state. Science. 2009;326:694-9 pubmed publisher..The stabilization of the mobile stalks of the ribosome also results in a more complete description of its structure. ..
- Laurberg M, Kristensen O, Martemyanov K, Gudkov A, Nagaev I, Hughes D, et al. Structure of a mutant EF-G reveals domain III and possibly the fusidic acid binding site. J Mol Biol. 2000;303:593-603 pubmed publisher..Possible locations for the effector loop and a possible binding site for fusidic acid are discussed in relation to some of the fusidic acid resistant mutations...
- Pavlov M, Antoun A, Lovmar M, Ehrenberg M. Complementary roles of initiation factor 1 and ribosome recycling factor in 70S ribosome splitting. EMBO J. 2008;27:1706-17 pubmed publisher..With support from our experimental data, we discuss the principally different mechanisms of ribosome splitting by IF1/IF3 and by RRF/EF-G. ..
- Nakamura Y. Molecular mimicry between protein and tRNA. J Mol Evol. 2001;53:282-9 pubmed..Structural, functional, and evolutionary aspects of molecular mimicry will be discussed. ..
- Rao A, Varshney U. Specific interaction between the ribosome recycling factor and the elongation factor G from Mycobacterium tuberculosis mediates peptidyl-tRNA release and ribosome recycling in Escherichia coli. EMBO J. 2001;20:2977-86 pubmed publisher..In addition, complementation of E.coli PEM100 (fusA(ts)) with MtuEFG suggests that the mechanism of RRF function is independent of the translocation activity of EFG...
- Valente L, Tiranti V, Marsano R, Malfatti E, Fernandez Vizarra E, Donnini C, et al. Infantile encephalopathy and defective mitochondrial DNA translation in patients with mutations of mitochondrial elongation factors EFG1 and EFTu. Am J Hum Genet. 2007;80:44-58 pubmed
- Liljas A. Imprinting through molecular mimicry. Protein synthesis. Curr Biol. 1996;6:247-9 pubmed..Part of the structure of translational elongation factor G, in a complex with GDP, resembles the tRNA bound in a ternary complex with elongation factor Tu and GTP; this 'molecular mimicry' extends to charge distribution as well as shape. ..
- 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. ..