Genetic Engineering of Bacterial and Eukaryotic Ribosomal Proteins for Investigation on Elongation Arrest of Nascent Polypeptides and Cell Differentiation

  • Fotini Leontiadou
  • Christina Matragkou
  • Filippos Kottakis
  • Dimitrios L. Kalpaxis
  • Ioannis S. Vizirianakis
  • Sofia Kouidou
  • Asterios S. Tsiftsoglou
  • Theodora Choli-Papadopoulou
Part of the Principles and Practice book series (PRINCIPLES)


This paper describes our efforts to investigate the role of specific bacterial and eukaryotic ribosomal proteins in crucial cell functions such as elongation arrest of the nascent polypeptides and cell differentiation. These objectives have been approached by: (1) engineering the L4 bacterial ribosomal protein, which has been shown by crystallographic data to be a candidate molecule for controlling the nascent polypeptides before their emerge from the ribosome, to elucidate specific amino-acids functions in highly conserved regions; and (2) isolating specific eukaryotic genes, like S5 and L35a, that encode ribosomal proteins to study their possible involvement in hematopoietic cell differentiation and apoptosis.


Ribosomal Protein K562 Cell Ribosomal Protein Gene Nascent Polypeptide Large Ribosomal Subunit 
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  1. Bessis M, Lessin LS and Beutler E (1983) Morphology of the erythron. In: Williams, WJ, Beutler E, Erslev AJ, Lichtman MA (eds) Hematology. New York: McGraw-Hill, pp 257–279Google Scholar
  2. Chittum HS and Champney WS (1994) Ribosomal protein gene sequence changes in erythromycin-resistance mutants of Escherichia coli. J. Bacteriol. 176: 6192–6198PubMedGoogle Scholar
  3. Ferrari S, Afredini R, Tagliafico E, Rossi E, Donelli A, Torelli G and Torelli U (1990). Non-coordinated expression of S6, S11, and S14 ribosomal protein genes in leukemic blast cells. Cancer Res. 50: 5825–5828PubMedGoogle Scholar
  4. Gabashvili IS, Agrawal RK, Spahn CM, Grassucci RA, Svergun DI, Frank J and Penezek P (2000) Solution structure of the E. coli 70S ribosome at 11.5 A resolution. Cell 100: 537–549PubMedCrossRefGoogle Scholar
  5. Hensold JO, Barth-Baus D and Stratton CA (1996) Inducers of erythroleukemic differentiation cause messenger RNAs that lack poly(A)-binding protein to accumulate in translationally inactive, salt-labile 80 S ribosomal complexes. J. Biol. Chem. 271, 23246–23254Google Scholar
  6. Herold M and Nierhaus KH (1987) Incorporation of six additional proteins to complete the assemply map of the 50S subunit from Escherichia coli ribosomes. J.Biol.Chem. 262: 8826–8833PubMedGoogle Scholar
  7. Kajikawa S, Nakayama H, Suzuki M, Takashima A, Murayama O, Nishihara M, Takahashi M and Doi K (1998) Increased expression of rat ribosomal protein L4 mRNA in 5-azacytidine-treated PC12 cells prior to apoptosis. Biochem. Biophys.Res. Commun. 252: 220–224Google Scholar
  8. Katunin VI, Muth GW, Strobel SA, Wintermeyer W and Rodnina MV (2002) Important contribution to catalysis of peptide Bond Formation by a single ionizing group within the ribosome. Mol. Cell 10: 339–346Google Scholar
  9. Lin CH, Palma JF and Solomon WB (1994) Phorbol ester induction of differentiation and apoptosis in the K562 cell line is accompanied by marked decreases in the stability of globin mRNAs and decreases in the steady state level of mRNAs encoding for ribosomal proteins L35, L31, L27, and L21. Cell. Mol. Biol. Res. 40: 13–26Google Scholar
  10. Lovett PS and Rogers EJ (1996). Translation attenuation regulation of chloramphenicol resistance in bacteria-a review. Microbiol. Rev. 60: 366–385PubMedGoogle Scholar
  11. Maheshwari Y, Rao M, Sykes DE, Tyner AL and Weiser MM (1993) Changes in ribosomal protein and ribosomal RNA synthesis during rat intestinal differentiation. Cell Growth Differ. 4: 745–752PubMedGoogle Scholar
  12. Mailhammer R, Szots H, Bonisch J and Dormer P (1992) Downregulation of messenger RNA levels for ribosomal proteins in differentiating HL-60 cells. Exp. Cell Res. 200: 145–148Google Scholar
  13. Marks PA, Sheffery M, Rifkind RA (1987) Induction of transformed cells to terminal differentiation and the modulation of gene expression. Cancer Res. 47: 659–666PubMedGoogle Scholar
  14. Milligan RA and Unwin PN (1986) Location of exit channel for nascent protein in 80S ribosome. Nature 319: 693–695PubMedCrossRefGoogle Scholar
  15. Naora H, Takai I, Adachi M and Naora H (1998) Altered cellular responses by varying expression of a ribosomal protein gene: Sequential coordination of enhancement and suppression of ribosomal protein S3a gene expression induces apoptosis. J. Cell Biol., 141: 741–753PubMedCrossRefGoogle Scholar
  16. Nissen P, Hansen J, Ban N, Moore PB, and Steitz TA (2000) The structural basis of ribosome activity in peptide bond synthesis. Science 289: 920–930PubMedCrossRefGoogle Scholar
  17. Pappas IS, Vizirianakis IS, Tsiftsoglou AS (2001) Cloning, sequencing and expression of a cDNA encoding the mouse L35a ribosomal protein during differentiation of murine erythroleukemia (MEL) cells. Cell Biol. Int. 25: 629–634Google Scholar
  18. Picking WD, Picking WL, Odom OW and Hardesty B (1992) Fluorescence characterization of the environment encountered by nascent polyalanine and polyserine as they exit Escherichia coli ribosomes during translation. Biochemistry 31: 2368–2375PubMedCrossRefGoogle Scholar
  19. Pool MR, Stumm J, Fulga TA, Sinning I and Dobberstein B (2002) Distinct modes of signal recognition particle interaction with the ribosome. Science 297: 1345–1348PubMedCrossRefGoogle Scholar
  20. Purohit P and Stern S (1994) Interactions of a small RNA with antibiotic and RNA ligands of the 30S subunit. Nature 370: 659–662PubMedCrossRefGoogle Scholar
  21. Schroeder R (1994) Translation: Dissecting RNA function. Nature 370: 597–598PubMedCrossRefGoogle Scholar
  22. Stelzl U, Spahn CMT and Nierhaus KH (2000) Selecting rRNA binding sites for the ribosomal proteins L4 and L6 from randomly fragmented rRNA: application of a method called SERF. Proc. Natl. Acad. Sci. USA 97: 4597–4602Google Scholar
  23. Su S and Bird RC (1995) Cell cycle, differentiation and tissue-independent expression of ribosomal protein L37. Eur. J. Biochem. 232: 789–797PubMedCrossRefGoogle Scholar
  24. Tenson T. and Ehrenberg M. (2002) Regulatory Nascent Peptides in the ribosomal tunnel. Cell 108: 591–594PubMedCrossRefGoogle Scholar
  25. Todorov IT, Noll F and Hadjiolov AA (1983) The sequential addition of ribosomal proteins during the formation of the small ribosomal subunit in Friend erythroleukemia cells. Eur. J. Biochem. 131, 271–275Google Scholar
  26. Tsiftsoglou AS, Wong W, Volloch V, Gusella J, Housman D (1982) Commitment of murine erythroleukemia (MEL) cells to terminal differentiation is associated with coordinated expression of globin and ribosomal genes. Prog. Clin. Biol. Res. 102A: 69–79Google Scholar
  27. Tsiftsoglou AS and Wong W (1985) Molecular and cellular mechanisms of leukemic hematopoietic cell differentiation: An analysis of the Friend system. Anticancer Res. 5: 81–100PubMedGoogle Scholar
  28. Tsiftsoglou AS, Hensold J, Robinson SH, Wong W (1987) The regulatory role of commitment in gene expression during induction of leukemic cell differentiation. In: Harrap KR, Connors TA (eds) New Avenues in Developmental Cancer Chemotherapy. Academic Press, New York, pp 205–227Nature Rev. Genet. 1: 57–64Google Scholar
  29. Tsiftsoglou AS, Robinson SH (1992) Differentiation of murine erythroleukemia cells and human HL-60 cell lines. In: Murphy MJ, Jr (eds) Concise Reviews in Clinical and Experimental Hematology. AlphaMed Press, Dayton, Ohio, pp 295–306Google Scholar
  30. Tsiftsoglou AS, Pappas IS and Vizirianakis IS (2002) The developmental program of murine erythroleukemia cells. Oncol. Res. 13: 339–349Google Scholar
  31. Unge J, Aberg A, Al-Kharadaghi S, Nikulin A, Nikonov S, Davydova NL, Nevskaya N, Garber M and Liljas A (1998) The crystal structure of the ribosomal protein L22 from Thermus thermophilus: insights into the mechanism of erythromycin resistance. Structure 6: 1577–1586PubMedCrossRefGoogle Scholar
  32. Vazquez D (1975) Inhibitors of Protein Synthesis. Springer, BerlinGoogle Scholar
  33. Vizirianakis IS, Pappas IS, Gougoumas D, Tsiftsoglou AS (1999) Expression of ribosomal protein S5 cloned gene during differentiation and apoptosis in murine erythroleukemia (MEL) cells. Oncol. Res., 11: 409–419Google Scholar
  34. Webb Y, Zhou X, Ngo L, Cornish V, Stahl J, Erdjument-Bromage H, Tempst P, Rifkind RA, Marks PA, Breslow A and Richon VM (1999) Photoaffinity labeling and mass spectrometry identify ribosomal protein S3 as a potential target for hybrid polar cytodifferentiation agents. J. Biol. Chem. 274: 14280–14287Google Scholar
  35. Wool IW (1993) The bifunctional nature of ribosomal proteins and speculations on their origins. In Nierhaus KH, Franceschi F, Subramania AR, Erdmann VA and WittmannLiebold B (eds) The Translational Apparatus: Structure, Function, Regulation, Evolution. Plenum Press, New York pp. 727–737CrossRefGoogle Scholar
  36. Wool IG, Chan YL and Gluck A (1995) Structure and evolution of mammalian ribosomal proteins. Biochem. Cell Biol. 73, 933–947Google Scholar
  37. Xu L, He GP, Li A and Ro HS (1994) Molecular characterization of the mouse ribosomal protein S24 multigene family: a uniquely expressed intron-containing gene with cell-specific expression of three alternatively spliced mRNAs. Nucleic Acid Res. 22: 646–655PubMedCrossRefGoogle Scholar
  38. Yonath A, Leonard KR and Wittmann HG (1987) A tunnel in the large ribosomal subunit revealed by three-dimensional image reconstruction. Science 236: 813–816PubMedCrossRefGoogle Scholar
  39. Zubay G (1973) In vitro synthesis of protein in microbial systems. Annu. Rev. Genet. 7: 267–287PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2004

Authors and Affiliations

  • Fotini Leontiadou
  • Christina Matragkou
  • Filippos Kottakis
  • Dimitrios L. Kalpaxis
  • Ioannis S. Vizirianakis
  • Sofia Kouidou
  • Asterios S. Tsiftsoglou
  • Theodora Choli-Papadopoulou

There are no affiliations available

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