Biochemistry (Moscow)

, Volume 78, Issue 12, pp 1354–1357 | Cite as

Internal initiation of polyuridylic acid translation in bacterial cell-free system

  • E. A. Sogorin
  • S. Ch. Agalarov
  • A. S. SpirinEmail author


The task of the present work was to answer the question: is the free 5′-end needed for effective translation of a model polyribonucleotide template — polyuridylic acid — in a bacterial (E. coli) cell-free system? For this purpose, the template activities of the original polyuridylic acid with its free 5′-end and the polyuridylic acid with blocked 5′-end were compared in the bacterial cell-free translation system. To block the 5′-end, the cytidylic oligodeoxyribonucleotide with fluorescein residue at its 5′-end and uridylic oligoribonucleotide sequence at its 3′-end, schematically described as FAM(dC)10(rU)50, was covalently attached (ligated) to the 5′-end of the template polyuridylic acid. It was shown that the efficiency of polyphenylalanine synthesis on the 5′-blocked template and on the polyuridylic acid with free 5′-end was virtually the same. It was concluded that bacterial ribosomes are capable of effectively initiating translation at the polyuridylic sequence independently of the 5′-end of template polyribonucleotide, i.e. via an internal initiation mechanism, in the absence of a Shine-Dalgarno sequence and AUG start codon.

Key words

polyuridylic acid translation initiation polyphenylalanine synthesis T4 RNA ligase cell-free translation 





N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid


polyuridylic acid


Tris-borate buffer


trichloroacetic acid




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Copyright information

© Pleiades Publishing, Ltd. 2013

Authors and Affiliations

  • E. A. Sogorin
    • 1
  • S. Ch. Agalarov
    • 1
  • A. S. Spirin
    • 1
    Email author
  1. 1.Institute of Protein ResearchRussian Academy of SciencesPushchino, Moscow RegionRussia

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