Abstract
EF-Tu fromE. coli, one of the superfamily of GTPase switch proteins, plays a central role in the fast and accurate delivery of aminoacyl-tRNAs to the translating ribosome. An overview is given about the regulatory effects of methylation, phosphorlation and phage-induced cleavage of EF-Tu on its function. During exponential growth, EF-Tu becomes monomethylated at Lys56 which is converted to Me2Lys upon entering the stationary phase. Lys56 is in the GTPase switch-1 regions (residues 49–62), a strongly conserved site involved in interactions with the nucleotide and the 5′ end of tRNA. Methylation was found to attenuate GTP hydrolysis and may thus enhance translational accuracy.In vivo 5–10% of EF-Tu is phosphorylated at Thr382 by a ribosome-associated kinase. In EF-Tu-GTP, Thr382 in domain 3 has a strategic position in the interface with domain 1; it is hydrogen-bonded to Glu117 that takes part in the switch-2 mechanism, and is close to the T-stem binding site of the tRNA, in a region known for many kirromycin-resistance mutations. Phosphorylation is enhanced by EF-Ts, but inhibited by kirromycin. In reverse, phosphorylated EF-Tu has an increased affinity for EF-Ts, does not bind kirromycin and can no longer bind aminoacyl tRNA. Thein vivo role of this reversibles modification is still a matter of speculation. T4 infection ofE. coli may trigger a phage-exclusion mechanism by activation of Lit, a host-encoded proteinase. As a result, EF-Tu is cleaved site-specifically between Gly59-Ile60 in the switch-1 region. Translation was found to drop beyond a minimum level. Interestingly, the identical sequence in the related EF-G appeared to remain fully intact. Although the Lit cleavage-mechanism may eventually lead to programmed cell death, the very efficient prevention of phage multiplication may be caused by a novel mechanisms ofin cis inhibition of late T4 mRNA translation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abel K., Jurnak F.: A complex profile of protein elongation: translating chemical energy into molecular movement.Structure4, 229–238 (1996b).
Abel K., Yoder M.D., Hilgenfeld R., Jurnak F.: An α to β conformational switch in EF-Tu.Structure4, 1153–1159 (1996a).
Aletta J.M., Cimato T.R., Ettinger M.J.: Protein methylation: a signal event in post-translational modification.Trends Biochem. Sci.23, 89–91 (1998).
Alexander C., Bilgin N., Lindschau C., Mesters J.R., Kraal B., Hilgenfeld R., Erdmann V.A., Lippmann C.: Phosphorylation of elongation factur Tu prevents ternary complex formation.J. Biol. Chem.270, 14541–14547 (1995).
Arai K., Clark B.F.C., Duffy L., Jones M.D., Kaziro Y., Laursen R.A., I'Italien J., Miller D.L., Nagarkatti S., Nakamura S., Nielsen K.M., Petesen T.E., Takayashi K., Wade M.: Primary structure of elongation factor Tu fromEscherichia coli.Proc. Nat. Acad. Sci. USA77, 1326–1330 (1980).
Berchtold H., Reshetnikova L., Reiser C.O.A., Schirmer N.K., Sprinzl M., Hilgenfeld R.: Crystal structure of active elongation factor Tu reveals major domain rearrangements.Nature365, 126–132 (1993).
Blumenthal T., Douglass J., Smith D.: Conformational alteration of protein synthesis elongation factor FE-Tu by EF-Ts and kirromycin.Proc. Nat. Acad. Sci. USA74, 3264–3267 (1977).
Bosch L., Kraal B., Van der Meide P.H., Duisterwinkel F.J., Van Noort J.M.: The elongation factor EF-Tu and its two encoding genes.Progr. Nucl. Acids Res. Molec. Biol.30, 91–126 (1983).
Bourne H.R., Sanders D.A., Mccormick F.: The GTPase superfamily: conserved structure and molecular mechanism.Nature349, 117–127 (1991).
Caldas T.D., El Yaagoubi A., Richarmer G.: Chaperone properties of bacterial elongation factor EF-Tu.J. Biol. Chem.273, 11478–11482 (1998).
Chang Y.W.E., Traugh J.A.: Phosphorylation of elongation factor 1 and ribosomal protein S6 by multipotential S6 kinase and insulin stimulation of translational elongation.J. Biol. Chem.272, 28252–28257 (1997).
Chang Y.W.E., Traugh J.A.: Insulin stimulation of phosphorylation of elongation factor 1 (eEF-1) enhances elongation activity.Eur. J. Biochem.251, 201–207 (1998).
Dever T.E., Costello C.E., Owens C.L., Rosenberry T.L., Merrick W.C.: Location of seven post-translational modifications in rabbit elongation factor 1α including dimethyllysine, trimethyllysine, and glycerylphosphorylethanolamine.J. Biol. Chem.264, 20518–20525 (1989).
Georgiou T., Yu Y.T.N., Ekunwe S., Buttner M.J., Zuurmond A.M., Kraal B., Kleanthous C., Snyder L.: Specific peptideactivated proteolytic cleavage ofEscherichia coli elongation factor Tu.Proc. Nat. Acad. Sci. USA95, 2891–2895 (1998).
Hughes S.M.: Are guanine nucleotide binding proteins a distinct class of regulatory proteins?.FEBS Lett.164, 1–8 (1983).
Kawashima T., Berthetcolominas C., Wulff M., Cusack S., Leberman R.: The structure of theEscherichia coli EF-Tu:EF-Ts complex at 2.5 ångstrom resolution.Nature379, 511–518 (1996).
Kjeldgaard M., Nissen P., Thirup S., Nyborg J.: The crystal structure of elongation factor EF-Tu fromThermus aquaticus in the GTP conformation.Structure1, 35–50 (1993).
Kraal B., Zeff L.A.H., Mesters J.R., Boon K., Vorstenbosch E.L.H., Bosch L., Anborgh P.H., Parmeggiani A., Hilgenfeld R.: Antibiotic resistance mechanisms of mutant EF-Tu species inEscherichia coli.Biochem. Cell. Biol.73, 1167–1177 (1995).
Kudlicki W., Coffman A., Kramer G., Hardesty B.: Renaturation of rhodanese by translational elongation factor EF-Tu. Protein refolding by EF-Tu flexing.J. Biol. Chem.272, 32206–32209 (1997).
Mesters J.R., Zeef L.A.H., Hilgenfeld R., de Graaf J.M., Kraal L., Bosch L.: The structural and functional basis for the kirromycin resistance of mutant EF-Tu species inEscherichia coli.EMBO J.13, 4877–4885 (1994).
Lippmann C., Lindschau C., Vijgenboom E., Schroder W., Bosch L., Erdmann V.A.: Prokaryotic elongation factor Tu is phosphorylatedin vivo.J. Biol. Chem.268, 601–607 (1993).
Mukulík K., Janda I.: Protein kinase associated with ribosomes phosphorylates ribosomal proteins ofStreptomyces collinus.Biochem. Biophys. Res. Commun.238, 370–376 (1997).
Nissen P., Kjeldgaard M., Thirup S., Polekhina G., Reshetnikova L., Clark B.F.C., Nyborg J.: Crystal structure of the ternary complex of Phe-tRNAPhe, EF-Tu, and a GTP analog.Science270, 1464–1472 (1995).
Nissen P., Kjeldgaard M., Thirup S., Clark B.F.C., Nyborg J.: The ternary complex of aminoacylated tRNA and EF-Tu GTP: recognition of a bond and a fold.Biochimie78, 921–933 (1996).
Nygård O., Nilsson L.: Translational dynamics. Interactions between the translational factors, tRNA and ribosomes during eukaryotic protein synthesis.Eur. J. Biochem.191, 1–17 (1990).
Parmeggiani A., Swart G.W.M.: Mechanism of action of kirromycin-like antibiotics.Ann. Rev. Microbiol.39, 557–577 (1985).
Peters H.I., Chang Y.W.E., Traugh J.A.: Phosphorylation of elongation factor 1 (EF-1) by protein kinase C stimulates GDP/GTP-exchange activity.Eur. J. Biochem.234, 550–556 (1995).
Plath T., Knudsen C., Bilgin N., Lindschau C., Erdmann V., Lippmann C.: Threonine 382 is essential for EF-Tu function, pp. 9–10 inAbstr. Book 17th Internat. tRNA Workshop, Chiba (Japan) 1997.
Polekhina G., Thirup S., Kjeldgaard M., Nissen P., Lippmann C., Nyborg J.: Helix unwinding in the effector region of elongation factor EF-Tu-GDP.Structure4, 1141–1151 (1996).
Robertson E.S., Aggison L.A., Nicholson A.W.: Phosphorylation of elongation factor G and ribosomal protein S6 in bacteriophage T7-infectedEscherichia coli.Mol. Microbiol.11, 1045–1057 (1994).
Van der Meide P.H., Vijgenboom E., Dicke M., Bosch L.: Regulation of the expression oftufA andtufB, the two genes coding for the elongation factor EF-Tu inEscherichia coli.FEBS Lett.139, 325–330 (1982).
Van Noort J.M., Kraal B., Sinjorgo K.M.C., Persoon N.L.M., Johanns E.S.D., Bosch L.: Methylationin vivo of elongation factor EF-Tu at lysine-56 decreases the rate of tRNA-dependent GTP hydrolysis.Eur. J. Biochem.160, 557–561 (1986).
Vorstenbosch E., Pape T., Rodnina M.V., Kraal B., Wintermeyer W.: The G222D mutation in elongation factor Tu inhibits the codon-induced conformational changes leading to GTPase activation on the ribosome.EMBO J.15, 6766–6774 (1996).
Wittinghofer A., Frank R., Leberman R.: Composition and properties of trypsin-cleaved elongation factor Tu.Eur. J. Biochem.108, 423–431 (1980).
Young C.C., Bernlohr R.W.: Elongation factor Tu is methylated in response to nutrient deprivation inEscherichia coli.J. Bacteriol.173, 3096–3100 (1991).
Yu Y.T.N., Snyder L.: Translation elongation factor Tu cleaved by a phage-exclusion system.Proc. Nat. Acad. Sci. USA91, 802–806 (1994).
Zeidler W., Schirmer N.K., Egle C., Ribeiro S., Kreutzer R., Sprinzl M.: Proteolysis and amino acid replacements in the effector region ofThermus thermophilus elongation factor Tu.Eur. J. Biochem.239, 265–271 (1996).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kraal, B., Lippmann, C. & Kleanthous, C. Translational regulation by modifications of the elongation factor Tu. Folia Microbiol 44, 131–141 (1999). https://doi.org/10.1007/BF02816232
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02816232