About this Journal Submit a Manuscript Table of Contents
BioMed Research International
Volume 2013 (2013), Article ID 585748, 12 pages
http://dx.doi.org/10.1155/2013/585748
Research Article

Cloning and Characterization of EF-Tu and EF-Ts from Pseudomonas aeruginosa

Chemistry Department, SCIE. 3.320, The University of Texas-Pan American, 1201 W. University Drive, Edinburg, TX 78541, USA

Received 1 May 2013; Accepted 12 July 2013

Academic Editor: Leonid Medved

Copyright © 2013 Stephanie O. Palmer et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Linked References

  1. G. Maschmeyer and I. Braveny, “Review of the incidence and prognosis of Pseudomonas aeruginosa infections in cancer patients in the 1990s,” European Journal of Clinical Microbiology and Infectious Diseases, vol. 19, no. 12, pp. 915–925, 2000. View at Publisher · View at Google Scholar · View at Scopus
  2. H. Giamarellou and A. Antoniadou, “Antipseudomonal antibiotics,” Medical Clinics of North America, vol. 85, no. 1, pp. 19–42, 2001. View at Publisher · View at Google Scholar · View at Scopus
  3. P. Roussel and G. Lamblin, “The glycosylation of airway mucins in cystic fibrosis and its relationship with lung infection by Pseudomonas aeruginosa,” Advances in Experimental Medicine and Biology, vol. 535, pp. 17–32, 2002. View at Scopus
  4. A. Oliver, R. Cantón, P. Campo, F. Baquero, and J. Blázquez, “High frequency of hypermutable Pseudomonas aeruginosa in cystic fibrosis lung infection,” Science, vol. 288, no. 5469, pp. 1251–1253, 2000. View at Publisher · View at Google Scholar · View at Scopus
  5. D. F. Sahm, D. C. Draghi, R. N. Master et al., “Pseudomonas aeruginosa antimicrobial resistance update: US resistance trends from 1998 to 2001,” p. 91, Proceedings of the 42nd Interscience Conference on Antimicrobial Agents and Chemotherapy (ICAAC '02), 2002.
  6. S. A. Nouer, M. Pinto, L. Teixeira, and M. Nucci, “Risk factors for multi-drug resistant Pseudomonas aeruginosa (MDRPa) colonization or infection in hospitalized patients,” in Proceedings of the 42nd Interscience Conference on Antimicrobial Agents and Chemotherapy (ICAAC '02), p. 340, 2002.
  7. D. J. C. Knowles, N. Foloppe, N. B. Matassova, and A. I. H. Murchie, “The bacterial ribosome, a promising focus for structure-based drug design,” Current Opinion in Pharmacology, vol. 2, no. 5, pp. 501–506, 2002. View at Publisher · View at Google Scholar · View at Scopus
  8. P. Schimmel, J. Tao, and J. Hill, “Aminoacyl tRNA synthetases as targets for new anti-infectives,” FASEB Journal, vol. 12, no. 15, pp. 1599–1609, 1998. View at Scopus
  9. R. C. Thompson, D. B. Dix, and A. M. Karim, “The reaction of ribosomes with elongation factor Tu.GTP complexes. Aminoacyl-tRNA-independent reactions in the elongation cycle determine the accuracy of protein synthesis,” Journal of Biological Chemistry, vol. 261, no. 11, pp. 4868–4874, 1986. View at Scopus
  10. M. Sprinzl, “Elongation factor Tu: a regulatory GTPase with an integrated effector,” Trends in Biochemical Sciences, vol. 19, no. 6, pp. 245–250, 1994. View at Publisher · View at Google Scholar · View at Scopus
  11. D. L. Miller and H. Weissbach, “Interactions between the elongation factors: the displacement of GDP from the Tu-GDP complex by factor Ts,” Biochemical and Biophysical Research Communications, vol. 38, no. 6, pp. 1016–1022, 1970. View at Scopus
  12. W. Ribble, W. E. Hill, U. A. Ochsner et al., “Discovery and analysis of 4H-Pyridopyrimidines, a class of selective bacterial protein synthesis inhibitors,” Antimicrobial Agents and Chemotherapy, vol. 54, no. 11, pp. 4648–4657, 2010. View at Publisher · View at Google Scholar · View at Scopus
  13. J. A. Leeds, M. J. LaMarche, J. T. Brewer et al., “In vitro and in vivo activities of novel, semisynthetic thiopeptide inhibitors of bacterial elongation factor Tu,” Antimicrobial Agents and Chemotherapy, vol. 55, no. 11, pp. 5277–5283, 2011. View at Publisher · View at Google Scholar · View at Scopus
  14. M. M. K. Jayasekera, K. Onheiber, J. Keith et al., “Identification of novel inhibitors of bacterial translation elongation factors,” Antimicrobial Agents and Chemotherapy, vol. 49, no. 1, pp. 131–136, 2005. View at Publisher · View at Google Scholar · View at Scopus
  15. M. F. Tam, J. A. Dodd, and W. E. Hill, “Physical characteristics of 16 S rRNA under reconstitution conditions,” Journal of Biological Chemistry, vol. 256, no. 12, pp. 6430–6434, 1981. View at Scopus
  16. M. M. Bradford, “A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding,” Analytical Biochemistry, vol. 72, no. 1-2, pp. 248–254, 1976. View at Scopus
  17. M. G. Cull and C. S. McHenry, “Purification of Escherichia coli DNA polymerase III holoenzyme,” Methods in Enzymology, vol. 262, pp. 22–35, 1995. View at Publisher · View at Google Scholar · View at Scopus
  18. J. G. Grasselli and W. M. Ritchey, “Spectral data and physical constrants for organic compounds,” in CRC Atlas of Spectral Data and Physical Constants for Organic Compounds, pp. 387–390, CRC Press, New York, NY, USA, 1975.
  19. J. M. Bullard, Y.-C. Cai, Y. Zhang, and L. L. Spremulli, “Effects of domain exchanges between Escherichia coli and mammalian mitochondrial EF-Tu on interactions with guanine nucleotides, aminoacyl-tRNA and ribosomes,” Biochimica et Biophysica Acta, vol. 1446, no. 1-2, pp. 102–114, 1999. View at Publisher · View at Google Scholar · View at Scopus
  20. J. M. Ravel, R. L. Shorey, S. Froehner, and W. Shive, “A study of the enzymic transfer of aminoacyl-RNA to Escherichia coli ribosomes,” Archives of Biochemistry and Biophysics, vol. 125, no. 2, pp. 514–526, 1968. View at Scopus
  21. A. Louie and F. Jurnak, “Kinetic studies of Escherichia coli elongation factor Tu-guanosine 5′-triphosphate-aminoacyl-tRNA complexes,” Biochemistry, vol. 24, no. 23, pp. 6433–6439, 1985. View at Scopus
  22. A. Louie, N. S. Ribeiro, B. R. Reid, and F. Jurnak, “Relative affinities of all Escherichia coli aminoacyl-tRNAs for elongation factor Tu-GTP,” Journal of Biological Chemistry, vol. 259, no. 8, pp. 5010–5016, 1984. View at Scopus
  23. J. M. Bullard, J. C. Williams, W. K. Acker, C. Jacobi, N. Janjic, and C. S. McHenry, “DNA polymerase III holoenzyme from Thermus thermophilus identification, expression, purification of components, and use to reconstitute a processive replicase,” Journal of Biological Chemistry, vol. 277, no. 16, pp. 13401–13408, 2002. View at Publisher · View at Google Scholar · View at Scopus
  24. D. Kavaliauskas, P. Nissen, and C. R. Knudsen, “The busiest of all ribosomal assistants: elongation factor Tu,” Biochemistry, vol. 51, no. 13, pp. 2642–2651, 2012. View at Publisher · View at Google Scholar · View at Scopus
  25. I. M. Krab and A. Parmeggiani, “EF-Tu, a GTPase odyssey,” Biochimica et Biophysica Acta, vol. 1443, no. 1-2, pp. 1–22, 1998. View at Publisher · View at Google Scholar · View at Scopus
  26. T. Kawashima, C. Berthet-Colominas, M. Wulff, S. Cusack, and R. Leberman, “The structure of the Escherichia coli EF-Tu.EF-Ts complex at 2.5 Å resolution,” Nature, vol. 379, no. 6565, pp. 511–518, 1996.
  27. H. Berchtold, L. Reshetnikova, C. O. A. Reiser, N. K. Schirmer, M. Sprinzl, and R. Hilgenfeld, “Crystal structure of active elongation factor Tu reveals major domain rearrangements,” Nature, vol. 365, pp. 126–132, 1993.
  28. O. Wiborg, C. Andersen, C. R. Knudsen, T. J. Kristensen, and B. F. Clark, “Towards an understanding of structure-function relationships of elongation factor Tu,” Biotechnology and Applied Biochemistry, vol. 19, p. 1, 1994. View at Scopus
  29. P. Nissen, M. Kjeldgaard, S. Thirup et al., “Crystal structure of the ternary complex of Phe-tRNA(Phe), EF-Tu, and a GTP analog,” Science, vol. 270, no. 5241, pp. 1464–1472, 1995. View at Scopus
  30. Y. Jiang, S. Nock, M. Nesper, M. Sprinzl, and P. B. Sigler, “Structure and importance of the dimerization domain in elongation factor Ts from Thermus thermophilus,” Biochemistry, vol. 35, no. 32, pp. 10269–10278, 1996. View at Publisher · View at Google Scholar · View at Scopus
  31. M. G. Jeppesen, T. Navratil, L. L. Spremulli, and J. Nyborg, “Crystal structure of the bovine mitochondrial elongation factor Tu.Ts complex,” Journal of Biological Chemistry, vol. 280, no. 6, pp. 5071–5081, 2005. View at Publisher · View at Google Scholar · View at Scopus
  32. Y. Zhang, V. Sun, and L. L. Spremulli, “Role of domains in Escherichia coli and mammalian mitochondrial elongation factor ts in the interaction with elongation factor Tu,” Journal of Biological Chemistry, vol. 272, no. 35, pp. 21956–21963, 1997. View at Publisher · View at Google Scholar · View at Scopus
  33. D. L. Miller and H. Weissbach, “Studies on the purification and properties of factor Tu from E. coli,” Archives of Biochemistry and Biophysics, vol. 141, no. 1, pp. 26–37, 1970. View at Scopus
  34. D. L. Miller and H. Weissbach, “Elongation factor Tu and the aminoacyl-tRNA-EFTu-GTP complex,” Methods in Enzymology, vol. 30, pp. 219–232, 1974. View at Publisher · View at Google Scholar · View at Scopus
  35. Y.-C. Cai, J. M. Bullard, N. L. Thompson, and L. L. Spremulli, “Interaction of mammalian mitochondrial elongation factor EF-Tu with guanine nucleotides,” Protein Science, vol. 9, no. 9, pp. 1791–1800, 2000. View at Scopus
  36. H. Weissbach, B. Redfield, and N. Brot, “Further studies on the role of factors Ts and Tu in protein synthesis,” Archives of Biochemistry and Biophysics, vol. 144, no. 1, pp. 224–229, 1971. View at Scopus
  37. V. Chau, G. Romero, and R. L. Biltonen, “Kinetic studies on the interactions of Escherichia coli K12 elongation factor Tu with GDP and elongation factor Ts,” Journal of Biological Chemistry, vol. 256, no. 11, pp. 5591–5596, 1981. View at Scopus
  38. Y.-C. Cai, J. M. Bullard, N. L. Thompson, and L. L. Spremulli, “Interaction of mitochondrial elongation factor Tu with aminoacyl-tRNA and elongation factor Ts,” Journal of Biological Chemistry, vol. 275, no. 27, pp. 20308–20314, 2000. View at Publisher · View at Google Scholar · View at Scopus
  39. Y. W. Hwang and D. L. Miller, “A study of the kinetic mechanism of elongation factor Ts,” Journal of Biological Chemistry, vol. 260, no. 21, pp. 11498–11502, 1985. View at Scopus
  40. Y.-X. Zhang, J. Tao, M. Zhou et al., “Elongation factor Ts of Chlamydia trachomatis: structure of the gene and properties of the protein,” Archives of Biochemistry and Biophysics, vol. 344, no. 1, pp. 43–52, 1997. View at Publisher · View at Google Scholar · View at Scopus
  41. B. R. Bochner and B. N. Ames, “Complete analysis of cellular nucleotides by two-dimensional thin layer chromatography,” Journal of Biological Chemistry, vol. 257, no. 16, pp. 9759–9769, 1982. View at Scopus
  42. K. B. Gromadski, H.-J. Wieden, and M. V. Rodnina, “Kinetic mechanism of elongation factor Ts-catalyzed nucleotide exchange in elongation factor Tu,” Biochemistry, vol. 41, no. 1, pp. 162–169, 2002. View at Publisher · View at Google Scholar · View at Scopus
  43. K. L. Manchester, “Determination of the kinetics of guanine nucleotide exchange on EF-Tu and EF-Ts: continuing uncertainties,” Biochemical and Biophysical Research Communications, vol. 314, no. 1, pp. 1–5, 2004. View at Publisher · View at Google Scholar · View at Scopus
  44. F. Jurnak, S. Heffron, B. Schick, and K. Delaria, “Three-dimensional models of the GDP and GTP forms of the guanine nucleotide domain of Escherichia coli elongation factor Tu,” Biochimica et Biophysica Acta, vol. 1050, no. 1–3, pp. 209–214, 1990. View at Publisher · View at Google Scholar · View at Scopus
  45. A. Pingoud, C. Urbanke, G. Krauss, F. Peters, and G. Maass, “Ternary complex formation between elongation factor Tu, GTP and aminoacyl-tRNA: an equilibrium study,” European Journal of Biochemistry, vol. 78, no. 2, pp. 403–409, 1977. View at Scopus
  46. S. Nakamura, K. Arai, K. Takahashi, and Y. Kaziro, “Alignment of the tryptic fragments and location of sulfhydryl groups of the polypeptide chain elongation factor Tu,” Biochemical and Biophysical Research Communications, vol. 77, no. 4, pp. 1418–1424, 1977. View at Scopus
  47. H. Song, M. R. Parsons, S. Rowsell, G. Leonard, and S. E. V. Phillips, “Crystal structure of intact elongation factor EF-Tu from Escherichia coli in GDP conformation at 2.05 Å resolution,” Journal of Molecular Biology, vol. 285, no. 3, pp. 1245–1256, 1999. View at Publisher · View at Google Scholar · View at Scopus
  48. A. Wagner, I. Simon, M. Sprinzl, and R. S. Goody, “Interaction of guanosine nucleotides and their analogs with elongation factor Tu from Thermus thermophilus,” Biochemistry, vol. 34, no. 39, pp. 12535–12542, 1995. View at Scopus
  49. C. C. Hall, J. D. Watkins, and N. H. Georgopapadakou, “Comparison of the Tu elongation factors from Staphylococcus aureus and Escherichia coli: possible basis for elfamycin insensitivity,” Antimicrobial Agents and Chemotherapy, vol. 35, no. 11, pp. 2366–2370, 1991. View at Scopus
  50. C. J. Schwartzbach and L. L. Spremulli, “Bovine mitochondrial protein synthesis elongation factors. Identification and initial characterization of an elongation factor Tu-elongation factor Ts complex,” Journal of Biological Chemistry, vol. 264, no. 32, pp. 19125–19131, 1989. View at Scopus