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Free cytoplasmic messenger ribonucleoprotein complexes from rabbit reticulocytes

Molecular Biology Reports volume 5, pages 59–64 (1979)Cite this article

Abstract

Free cytoplasmic globin mRNA containing mRNP-particles were isolated from rabbit reticulocytes by zonal sucrose gradient centrifugation and their properties were compared with mRNP particles isolated in the same way from EDTA-dissociated reticulocyte polyribosomes. The average poly(A)-length of 9S mRNA from free cytoplasmic mRNP was 17–20 nucleotides being about two times shorter than the average poly(A)-length of polysomal 9S mRNA. The protein composition of the free cytoplasmic mRNP particles disclosed the absence of the 76,000 dalton protein which is associated with the 3′poly(A)-segment of polysomal globin mRNA. It was concluded that free cytoplasmic mRNP-particles from rabbit reticulocytes can be classified as “old” mRNP in a post-translational phase. Free cytoplasmic mRNPs were translated in heterologous cell-free systems as well as in Xenopus laevis oocytes. Addition of hemin stimulated the synthesis of α-globin in all systems, while the presence of the cap analogue m7G(5′)p inhibited translation of free cytoplasmic mRNA completely. The latter finding suggested that free cytoplasmic mRNA has a 5′ terminal “cap”. Shortening of the poly(A)-segment with concomitant loss of the 76,000 dalton protein may lead to less efficient translation of free cytoplasmic mRNP.

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References

  1. BlobelG., Proc. Natl. Acad. Sci. U.S.A. 70, 924 (1973).

    Google Scholar 

  2. SchwartzH. and DarnellJ.E., J. Mol. Biol. 104, 833 (1976).

    Google Scholar 

  3. JanssenD.B., Counotte-PotmanA.D. and vanVenrooijW.J., Mol. Biol. Rep. 3, 87 (1976).

    Google Scholar 

  4. VanVenrooijW.J., vanEekelenC.A.G., JansenR.T.P. and PrincenJ.M.G., Nature (Lond.) 270, 189 (1977).

    Google Scholar 

  5. PreobrazhenskiA.A. and SpirinA.S., Progress Nucl. Acid. Res. Mol. Biol. 21, 1 (1978).

    Google Scholar 

  6. SpirinA.S., Eur. J. Biochem. 10, 20 (1969).

    Google Scholar 

  7. SamarinaO.P., LukanidinE.M. and GeorgievG.P., Acta Endocrin. Suppl. 180, 143 (1973).

    Google Scholar 

  8. SlegersH. and KondoM., Nucleic Acid Res. 4, 625 (1977).

    Google Scholar 

  9. DworkinM.B. and InfanteA.A., Dev. Biol. 53, 73 (1976).

    Google Scholar 

  10. SonensheinG.E. and BrawermanG., Eur. J. Biochem. 73, 307 (1977).

    Google Scholar 

  11. YapS.H., StrairR.K. and ShafritzD.A., J. Biol. Chem. 253, 4944 (1978).

    Google Scholar 

  12. MarbaixG., HuezG., NokinP. and CleuterY. FEBS Letters 66, 269 (1976).

    Google Scholar 

  13. Bonanou-TzedakiS.A., PragnellI.B. and ArnsteinH.R.V. FEBS Letters 26, 77 (1972).

    Google Scholar 

  14. OlsenG.D., GaskillP. and KabatD. Biochim. Biophys. Acta 272, 297 (1972).

    Google Scholar 

  15. GianniA.M., GiglioniB., OttolenghiS., ComiP. and GuidottiG.G. Nature New Biol. 240, 183 (1972).

    Google Scholar 

  16. Jacobs-LorenaM. and BaglioniC. Proc. Natl. Acad. Sci. U.S.A. 69, 1425 (1972).

    Google Scholar 

  17. EfstratiadisA., KafatosF.C. and ManiatisT. Cell 10, 571 (1977).

    Google Scholar 

  18. Van-TanH. and SchapiraG. Eur. J. Biochem. 85, 271 (1978).

    Google Scholar 

  19. LaemmliU.K. Nature (Lond.) 227, 680 (1970).

    Google Scholar 

  20. GiglioniB., GianniA.M., ComiP., OttolenghiS. and RungyerD. Nature New Biol. 246, 99 (1973).

    Google Scholar 

  21. AsselbergsF.A.M., vanVenrooijW.J. and BloemendalH. Eur. J. Biochem. 87, 517 (1978).

    Google Scholar 

  22. ShatkinA.J. Cell, 9, 645 (1976).

    Google Scholar 

  23. HuntJ.A. and OakesG.N. Biochem. J. 155, 637 (1976).

    Google Scholar 

  24. MuthukrishnanS., BothG.W., FuruichiY. and ShatkinA.J. Nature 255, 33 (1975).

    Google Scholar 

  25. PerryR.P. and ScherrerK. FEBS Letters 57, 73 (1975).

    Google Scholar 

  26. ShafritzD.A., WeinsteinJ.A., SaferB., MerrickW.C., WeberL.A., HickeyE.D. and BaglioniC. Nature 261, 291 (1976).

    Google Scholar 

  27. HickeyE.D., WeberL.A. and BaglioniC. Proc. Natl. Acad. Sci. U.S.A. 73, 19 (1976).

    Google Scholar 

  28. SenG.C., LebleuB., BrownG.E., RebelloM.A. FuruichiY., MorganM., ShatkinA.J. and LengyelP. Biochem. Biophys. Res. Commun. 65, 427 (1975).

    Google Scholar 

  29. PelhamH.R. and JacksonR.I. Eur. J. Biochem. 67, 247 (1976).

    Google Scholar 

  30. MillerR.L. and SchweetR. Arch. Biochem. Biophys. 125, 632 (1968).

    Google Scholar 

  31. PhilipsJ.A., SnijderP.G. and KazazianH.H. Nature 269, 442 (1977).

    Google Scholar 

  32. SheinessD. and DarnellJ.E. Nature New Biol. 241, 265 (1973).

    Google Scholar 

  33. NokinP., HuezG., MarbaixG., BurnyA. and ChantrenneH. Eur. J. Biochem. 62, 509 (1976).

    Google Scholar 

  34. BrawermanG. Mol. Biol. Rep. 1, 7 (1973).

    Google Scholar 

  35. GorskiJ., MorrisonM.R., MerkelC.G. and LingrelJ.B. Nature 253, 749 (1975).

    Google Scholar 

  36. SoreqH., NudelU., SalomonR., RevelM. and LittauerU.Z. J. Mol. Biol. 88, 233 (1974).

    Google Scholar 

  37. SippelA.E., StavrianopoulosJ.G., SchutzG. and FeigelsonP. Proc. Natl. Acad. Sci. U.S.A. 71, 4635 (1974).

    Google Scholar 

  38. HuezG., MarbaixG., HubertE., CleuterY., LeclercqM., ChantrenneH., DevosR., SoreqH., NudelU. and LittauerU.Z., Eur. J. Biochem. 59, 589 (1975).

    Google Scholar 

  39. CivelliO., VincentA., BuriJ.F. and ScherrerK., FEBS Letters 72, 71 (1976).

    Google Scholar 

  40. LondonI.M., ClemensM.J., RanuR.S., LevinD.H., CherbasL.F. and ErnstV. Fed. Proc. 35, 2218 (1976).

    Google Scholar 

  41. GrossM. Arch. Biochem. Biophys. 180, 121 (1977).

    Google Scholar 

  42. MarbaixG., BurnyA., HuezG., LebleuB. and TemmermanJ. Eur. J. Biochem. 13, 322 (1970).

    Google Scholar 

  43. HuntR.T., HunterA.R. and MunroA.J. Nature 220, 481 (1968).

    Google Scholar 

  44. LodishH.F. and JacobsenM. J. Biol. Chem. 247, 3622 (1972).

    Google Scholar 

  45. LodishH.F. Nature 251, 385 (1974).

    Google Scholar 

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Affiliations

  1. Department of Biochemistry, University of Nijmegen, Geert Grooteplein Noord 21, Nijmegen, The Netherlands

    Hans M. G. Princen, Chris A. G. van Eekelen, Fred A. M. Asselbergs & Walther J. van Venrooij

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  1. Hans M. G. Princen
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  2. Chris A. G. van Eekelen
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  3. Fred A. M. Asselbergs
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  4. Walther J. van Venrooij
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Princen, H.M.G., van Eekelen, C.A.G., Asselbergs, F.A.M. et al. Free cytoplasmic messenger ribonucleoprotein complexes from rabbit reticulocytes. Mol Biol Rep 5, 59–64 (1979). https://doi.org/10.1007/BF00777489

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  • DOI: https://doi.org/10.1007/BF00777489

Keywords

  • Hemin
  • Xenopus Laevis
  • Protein Composition
  • Sucrose Gradient
  • Efficient Translation