Abstract
In the elongation cycle of polypeptide synthesis, elongation factor Tu(EF- Tu) undergoes a sequential series of interactions with a number of low (GTP and GDP) and high molecular weight (aa-tRNA, ribosomes, and EF-Ts) components (for a review, see Miller and Weissbach, 1977; Kaziro, 1978). Following its property as a guanine nucleotide binding protein, EF-Tu displays a GTPase activity when, as a component of the ternary complex EF-Tu-GTP-aa-tRNA, it interacts with the ribosome-mRNA complex. This activity has also been observed in the absence of ribosomes with EF-Tu alone by simply increasing the concentration of monovalent cations, as well as by utilizing the antibiotic kirromycin whose effect first allowed the identification of a catalytic center for GTP hydrolysis on the factor (for a review, Parmeggiani and Sander, 1980, 1981; Parmeggiani and Swart, 1985). In the past few years, the progress of our knowledge of the EF-Tu functions proceeded with that of its primary structure, three- dimensional conformation, and genetic situation (for a review see Bosch et al., 1983). The isolation of EF-Tu mutant factors was made possible by the use of kirromycin. These accomplishments offer us the possibility of new approaches for the study of the function-structure relationships of this important protein. In this context, besides intensifying the study of the presently available EF-Tu mutants selected with the help of kirromycin, we have started a program aimed at modifying the tufA gene in positions that may code for single amino acids or segments of the polypeptide chain important for the different EF-Tu functions.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Bosch, L., Kraal, B., Van der Meide, P.H., Duisterwinkel, F.J., Van Noort, J.M. (1983). The elongation factor EF-Tu and its two encoding genes. Prog. Nucl. Acid Res. Mol. Biol. 30: 91–126.
Chinali, G., Wolf, H., Parmeggiani, A. (1977). Effect of kirromycin on elongation factor Tu. Location of the catalytic center for ribosome elongation factor Tu GTPase activity on the elongation factor. Eur. J. Biochem. 75: 55–65.
Clark, B.F.C., LaCour, T.F.M., Fontecilla-Camps, J., Morikawa, K., Nielsen, K.M., Nyborg, J., Rubin, J.R. (1982). FEBS Meeting on Cell Function and Differentiation, Vol. 3, Symp. 12, pp. 59–64.
Dente, L., Cesareni, G., Cortese, R. (1983). pEMBL: a new family of single stranded plasmids. Nucl. Acids Res. 11: 1645–1656.
Duisterwinkel, F.J., De Graaf, J.M., Kraal, B., Bosch, L. (1981a). A kirromycin resistant elongation factor Tu from Escherichia coli contains a threonine instead of alanine residue in position 375. FEBS Lett. 13: 89–93.
Duisterwinkel, F.J., De Graaf, J.M., Schretlen, P.J.M., Kraal, B., Bosch, L. (1981 b). A mutant elongation factor Tu which does not immobilize the ribosome upon binding of kirromycin. Eur. J. Biochem. 117: 7–12.
Duisterwinkel, F.J., Kraal, B., De Graaf, J.M., Talens, A., Bosch, L., Swart, G.W.M., Parmeggiani, A., LaCour, T.F.M., Nyborg, J., Clark, B.F.C. (1984). Specific alterations of the EF-Tu polypeptide chain considered in the light of its three-dimensional structure. EMBO J. 3: 113–120.
Fasano, O., Parmeggiani, A. (1981). Altered regulation of the guanosine 5′-triphos- phatase activity in a kirromycin resistant elongation factor Tu. Biochemistry 20: 1361–1366.
Fischer, E., Wolf, H., Hantke, K., Parmeggiani, A. (1977). Elongation factor Tu resistant to kirromycin in an Escherichia coli mutant altered in both tuf genes. Proc. Natl. Acad. Sci. USA 74: 4341–4345.
Guerrier-Takada, C., Johnson, A., Miller, D.L., Cole, P.E. (1981). Transfer RNA cross-linked to the elongation factor Tu subunit of Qβ replicase does not inhibit Qβ RNA replication. J. Biol. Chem. 256: 5840–5844.
Halliday, K.R. (1983). Regional homology in GTP-binding proto-oncogene products and elongation factors. J. Cycl. Nucleot. Prot. Phosphor. Res. 9: 435–448.
Ivell, R., Fasano, O., Crechet, J.B., Parmeggiani, A. (1981). Characterization of a kirromycin-resistant elongation factor Tu from Escherichia coli. Biochemistry 20: 1355–1361.
Johnson, A.E., Miller, D.L., Cantor, C.R. (1978). Functional covalent complex between elongation factor Tu and an analog of lysyl-tRNA. Proc. Natl. Acad. Sci. USA 75: 3075–3079.
Jonák, J., Petersen, T.E., Clark, B.F.C., Rychlik, I. (1982). 7V-tosyl-L-phenylalan- ylchloromethane reacts with cysteine-81 in the molecule of elongation factor Tu from Escherichia coli. FEBS Lett. 1150: 485–488.
Jonák, J., Smrt, J., Holý, A., Rychlík, I. (1980). Interaction of Escherichia coli EF-Tu GTP and EF-Tu-GDP with analogues of the 3′ terminus of aa-tRNA. Eur. J. Biochem. 105: 315–320.
Jurnak, F. (1985). Structure of the GDP domain of EF-Tu and location of the amino acids homologous to ras oncogene protein. Science 230: 32–36.
Kaziro, Y. (1978). The role of guanosine 5′-triphosphate in polypeptide chain elongation. Biochem. Biophys. Acta 505: 95–127.
LaCour, T.F.M., Nyborg, J., Thirup, S., Clark, B.F.C. (1985). Structural details of the binding of guanosine diphosphate to elongation factor Tu from E. coli as studied by X-ray crystallography. EMBO J. 4: 2385–2388.
Leberman, R., Egner, U. (1984). Homologies in the primary structure of GTP- binding proteins: the nucleotide-binding site of EF-Tu and p21. EMBO J. 3: 339–341.
Miller, D.L., Weissbach, H. (1970). Interaction between the elongation factors: the displacement of GDP from the EF-Tu GDP complex by factor Ts. Biochem. Biophys. Res. Commun. 38: 1016–1022.
Miller, D.L., Weissbach, H. (1977). Factors involved in the transfer of aminoacyl- tRNA to the ribosomes. In: Molecular mechanisms in protein biosynthesis, eds. Weissbach, H., Petska, S. Academic Press, New York, pp. 323–373.
Miyajima, A., Kaziro, Y. (1980). The expression of the cloned tufB gene in vivo. FEBS Lett. 2: 215–218.
Parmeggiani, A., Sander, G. (1980). Properties and action of kirromycin (Mocimycin) and related antibiotics. Top. Antibiot. Chem. 5: 159–221.
Parmeggiani, A., Sander, G. (1981). Properties and regulation of the GTPase activities of elongation factors Tu and G, and of initiation factor 2. Mol. Gen. Biochem. 31: 129–158.
Parmeggiani, A., Swart, G.W.M. (1985). Mechanism of action of kirromycin-like antibiotics. Ann. Rev. Microbiol 39: 557–577.
Rubin, J.R., Morikawa, K., Nyborg, J., LaCour, T.F.M., Clark, B.F.C., Miller, D.L. (1981). Structural features of the GDP binding site of elongation factor Tu from Escherichia coli as determined by X-ray diffraction. FEBS Lett. 129: 177–179.
Sacerdot, C., Dessen, P., Hershey, J.W.B., Plumbridge, J.A., Grunberg-Manago, M. (1984). Sequence of the initiation factor IF-2 gene: unusual protein features and homologies with elongation factors. Proc. Acad. Natl. Sci. USA 81: 7787–7791.
Swart, G.W.M., Kraal, B., Bosch, L., Parmeggiani, A. (1981). Allosteric changes of the guanine nucleotide site of elongation factor EF-Tu. A comparative study of two kirromycin-resistant EF-Tu species. FEBS Lett. 142: 101–106.
Swart, G.W.M., Merola, M., Guesnet, J., Parmeggiani, A. (1984). Studies on the function-structure relationships in the interaction of elongation factor Tu with guanosine nucleotides, aminoacyl-tRNA and kirromycin. In: Metabolism and enzymology of the nucleic acids including gene manipulations, vol. 5, eds. Zelinka, J., Balan, J. Publishing House of the Slovak Academy of Science, Bratislava, pp. 277–288.
Traboni, C., Ciliberto, G., Cortese, R. (1982). A novel method for site-directed mutagenesis: its application to a eukaryotic tRNAPro gene promo tor. EMBO J. 1: 415–420.
Van de Klundert, J.A.M., Den Turk, E., Borman, A.H., Van der Meide, P.H., Bosch, L. (1977). Isolation and characterization of a mocimycin resistant mutant of Escherichia coli with an altered elongation factor EF-Tu. FEBS Lett. 81: 303–307.
Van der Meide, P.H., Duisterwinkel, F.J., De Graaf, J.M., Kraal, B., Bosch, L. (1981). Molecular properties of two mutant species of the elongation factor Tu. Eur. J. Biochem. 117: 1–6.
Van der Meide, P.H., Kastelein, R.A., Vijgenboom, E., Bosch, L. (1983). tuf gene dosage effects on the intracellular concentration of EF-TuB. Eur. J. Biochem. 130: 409–417.
Van Hemert, F.J., Amons, R., Pluijms, W.J.M., Van Ormondt, H., Moller, W. (1984). The primary structure of elongation factor EF-1 from the brine shrimp Artemia. EMBO J. 3: 1109–1113.
Van Noort, J.M., Kraal, B., Bosch, L., LaCour, T.F.M., Nyborg, J., Clark, B.F.C. (1985). Cross-linking of tRNA at two different sites of EF-Tu. Proc. Natl. Acad. Sci. USA 81: 3969–3972.
Wierenga, R.K., Hoi, W.G.J. (1983). Predicted nucleotide-binding properties of p21 protein and its cancer-associated variant. Nature 302: 842–844.
Wittinghofer, A., Warren, W.F., Leberman, R. (1977). Structural requirements of the GDP binding site of elongation factor Tu. FEBS Lett. 75: 241–243.
Wolf, H., Chinali, G., Parmeggiani, A. (1977). Mechanism of the inhibition of protein synthesis by kirromycin. Role of elongation factor Tu and ribosomes. Eur. J. Biochem. 75: 67–75.
Zengel, J.M., Lindahl, L. (1982). A secondary promoter for elongation factor Tu synthesis in the str ribosomal protein operon of Escherichia coli. Mol. Gen. Gen. 185: 487–492.
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1986 Springer-Verlag New York Inc.
About this chapter
Cite this chapter
Parmeggiani, A. et al. (1986). Elongation Factor Tu Mutants and Site-Directed Mutagenesis of tufA . In: Hardesty, B., Kramer, G. (eds) Structure, Function, and Genetics of Ribosomes. Springer Series in Molecular Biology. Springer, New York, NY. https://doi.org/10.1007/978-1-4612-4884-2_39
Download citation
DOI: https://doi.org/10.1007/978-1-4612-4884-2_39
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-4612-9346-0
Online ISBN: 978-1-4612-4884-2
eBook Packages: Springer Book Archive