Molecular Biology

, Volume 52, Issue 1, pp 15–18 | Cite as

Stimulation of the Translation of Reporter mRNA in the Presence of Another mRNA in a Cell-Free System Based on Wheat Germ Extract

  • E. A. Sogorin
  • S. Ch. AgalarovEmail author
Molecular Cell Biology


The translation of uncapped mRNAs encoding luciferase and green fluorescent protein in a cellfree translation system based on wheat germ extract has been studied. It turned out that two simultaneously translated (in one tube) different templates in a certain range of concentrations not only do not compete, but mutually enhance each other's translation. It has been shown that the synthesis of luciferase in the presence of mRNA that encodes green fluorescent protein is much more effective than in the translation of only luciferase mRNA at the same concentration. Similarly, the efficiency of the synthesis of green fluorescent protein increases in the presence of the template that encodes luciferase. It follows that the total effect of the concurrent translation of two different mRNAs exceeds the sum of the effects of the translation of each mRNA separately.


translation uncapped mRNAs polyribosomes 



untranslated region


green fluorescent protein

Luc mRNA

luciferase mRNA


GFP that encodes mRNA


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Gallie D.R. 1991. The cap and poly (A) tail function synergistically to regulate mRNA translational efficiency. Genes Dev. 5, 2108–2116.CrossRefPubMedGoogle Scholar
  2. 2.
    Michel Y.M., Poncet D., Piron M., et al. 2000. Cap-Poly(A) synergy in mammalian cell-free extracts. Investigation of the requirements for poly(A)-mediated stimulation of translation initiation}. J. Biol. Chem. 275, 32268–32276.CrossRefPubMedGoogle Scholar
  3. 3.
    Wells S.E., Hillner P.E., Vale R.D., et al. 1998. Circularization of mRNA by eukaryotic translation initiation factors. Mol. Cell. 2, 135–140.CrossRefPubMedGoogle Scholar
  4. 4.
    Philipps G.R. 1965. Haemoglobin synthesis and polysomes in intact reticulocytes. Nature. 205, 567–570.CrossRefPubMedGoogle Scholar
  5. 5.
    Alekhina O.M., Vassilenko K.S., Spirin A.S. 2007. Translation of non-capped mRNAs in a eukaryotic cell-free system: Acceleration of initiation rate in the course of polysome formation. Nucleic Acids Res. 35, 6547–6559.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Kopeina G.S., Afonina Zh.A., Gromova K.V., et al. 2008. Step-wise formation of eukaryotic double-row polyribosomes and circular translation of polysomal mRNA. Nucleic Acids Res. 36, 2476–2488.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Sogorin E.A., Agalarov S.Ch., Spirin A.S. 2015. Formation of new polysomes on free mRNAs in a cell-free translation systems is accompanied by partial disassembly of previously formed polysomes. Biochemistry (Moscow). 80, 1327–1330.CrossRefGoogle Scholar
  8. 8.
    Shirokov V.A., Kommer A., Kolb V.A., Spirin A.S. 2007. Continuous-exchange protein-synthesizing systems. Methods Mol. Biol. 375, 19–55.PubMedGoogle Scholar
  9. 9.
    Kolb V.A., Makeyev E.V., Spirin A.S. 1994. Folding of firefly luciferase during translation in a cell-free system. EMBO J. 13, 3631–3637.PubMedPubMedCentralGoogle Scholar
  10. 10.
    Sogorin E.A., Agalarov S.C., Spirin A.S. 2016. Interpolysomal coupling of termination and initiation during translation in eukaryotic cell-free system. Sci.Rep. 6, 24518.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Pleiades Publishing, Inc. 2018

Authors and Affiliations

  1. 1.Institute of Protein ResearchRussian Academy of SciencesPushchino, Moscow oblastRussia

Personalised recommendations