IDEAS home Printed from https://ideas.repec.org/a/nat/nature/v430y2004i7000d10.1038_nature02712.html
   My bibliography  Save this article

Structural basis for template-independent RNA polymerization

Author

Listed:
  • Kozo Tomita

    (Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology
    University of Tokyo)

  • Shuya Fukai

    (Tokyo Institute of Technology)

  • Ryuichiro Ishitani

    (the University of Tokyo)

  • Takuya Ueda

    (University of Tokyo)

  • Nono Takeuchi

    (University of Tokyo)

  • Dmitry G. Vassylyev

    (RIKEN Harima Institute at SPring-8)

  • Osamu Nureki

    (Tokyo Institute of Technology
    PRESTO, JST, Honcho)

Abstract

The 3′-terminal CCA nucleotide sequence (positions 74–76) of transfer RNA is essential for amino acid attachment1 and interaction with the ribosome2,3,4 during protein synthesis. The CCA sequence is synthesized de novo and/or repaired by a template-independent RNA polymerase, ‘CCA-adding enzyme’, using CTP and ATP as substrates5. Despite structural and biochemical studies5,6,7,8, the mechanism by which the CCA-adding enzyme synthesizes the defined sequence without a nucleic acid template remains elusive. Here we present the crystal structure of Aquifex aeolicus CCA-adding enzyme, bound to a primer tRNA lacking the terminal adenosine and an incoming ATP analogue, at 2.8 Å resolution. The enzyme enfolds the acceptor T helix of the tRNA molecule. In the catalytic pocket, C75 is adjacent to ATP, and their base moieties are stacked. The complementary pocket for recognizing C74-C75 of tRNA forms a ‘protein template’ for the penultimate two nucleotides, mimicking the nucleotide template used by template-dependent polymerases. These results are supported by systematic analyses of mutants. Our structure represents the ‘pre-insertion’ stage of selecting the incoming nucleotide and provides the structural basis for the mechanism underlying template-independent RNA polymerization.

Suggested Citation

  • Kozo Tomita & Shuya Fukai & Ryuichiro Ishitani & Takuya Ueda & Nono Takeuchi & Dmitry G. Vassylyev & Osamu Nureki, 2004. "Structural basis for template-independent RNA polymerization," Nature, Nature, vol. 430(7000), pages 700-704, August.
    • Handle: RePEc:nat:nature:v:430:y:2004:i:7000:d:10.1038_nature02712
    DOI: 10.1038/nature02712
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/nature02712
    File Function: Abstract
    Download Restriction: Access to the full text of the articles in this series is restricted.
    File URL: https://libkey.io/10.1038/nature02712?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    As the access to this document is restricted, you may want to search for a different version of it.

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:nature:v:430:y:2004:i:7000:d:10.1038_nature02712. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    We have no bibliographic references for this item. You can help adding them by using this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.