1. Biochemistry and Chemical Biology
  2. Structural Biology and Molecular Biophysics

Structural insights into the assembly and polyA signal recognition mechanism of the human CPSF complex

  1. Marcello Clerici
  2. Marco Faini
  3. Ruedi Aebersold  Is a corresponding author
  4. Martin Jinek  Is a corresponding author
  1. University of Zurich, Switzerland
  2. Swiss Federal Institute of Technology, Switzerland
https://doi.org/10.7554/eLife.33111
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  1. Marcello Clerici
  2. Marco Faini
  3. Ruedi Aebersold
  4. Martin Jinek
(2017)
Structural insights into the assembly and polyA signal recognition mechanism of the human CPSF complex
eLife 6:e33111.
https://doi.org/10.7554/eLife.33111
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Abstract

3' polyadenylation is a key step in eukaryotic mRNA biogenesis. In mammalian cells, this process is dependent on the recognition of the hexanucleotide AAUAAA motif in the pre-mRNA polyadenylation signal by the Cleavage and Polyadenylation Specificity Factor (CPSF) complex. A core CPSF complex comprising CPSF160, WDR33, CPSF30 and Fip1 is sufficient for AAUAAA motif recognition, yet the molecular interactions underpinning its assembly and mechanism of PAS recognition are not understood. Based on cross-linking-coupled mass spectrometry, crystal structure of the CPSF160-WDR33 subcomplex and biochemical assays, we define the molecular architecture of the core human CPSF complex, identifying specific domains involved in inter-subunit interactions. In addition to zinc finger domains in CPSF30, we identify using quantitative RNA binding assays an N-terminal lysine/arginine-rich motif in WDR33 as a critical determinant of specific AAUAAA motif recognition. Together, these results shed light on the function of CPSF in mediating PAS-dependent RNA cleavage and polyadenylation.

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Author details

  1. Marcello Clerici

    Department of Biochemistry, University of Zurich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.

  2. Marco Faini

    Department of Biology, Institute of Molecular Systems Biology, Swiss Federal Institute of Technology, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.

  3. Ruedi Aebersold

    Department of Biology, Institute of Molecular Systems Biology, Swiss Federal Institute of Technology, Zurich, Switzerland
    For correspondence
    aebersold@imsb.biol.ethz.ch
    Competing interests
    The authors declare that no competing interests exist.

  4. Martin Jinek

    Department of Biochemistry, University of Zurich, Zurich, Switzerland
    For correspondence
    jinek@bioc.uzh.ch
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7601-210X

Funding

European Research Council (ERC-StG-337284)

  • Marcello Clerici
  • Martin Jinek

European Molecular Biology Organization (ALTF-343-2013)

  • Marco Faini

European Research Council (HEALTH-F4-2008-201648)

  • Ruedi Aebersold

European Research Council (233226)

  • Ruedi Aebersold

H2020 European Research Council (670821)

  • Ruedi Aebersold

Innovative Medicines Initiative Joint Undertaking (ULTRA-DD grant no. 115766)

  • Ruedi Aebersold

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2017, Clerici et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Marcello Clerici
  2. Marco Faini
  3. Ruedi Aebersold
  4. Martin Jinek
(2017)
Structural insights into the assembly and polyA signal recognition mechanism of the human CPSF complex
eLife 6:e33111.
https://doi.org/10.7554/eLife.33111

Share this article

https://doi.org/10.7554/eLife.33111

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