1. Chromosomes and Gene Expression
  2. Structural Biology and Molecular Biophysics

Fip1 is a multivalent interaction scaffold for processing factors in human mRNA 3' end biogenesis

  1. Lena Maria Muckenfuss
  2. Anabel Carmen Migenda Herranz
  3. Franziska M Boneberg
  4. Marcello Clerici
  5. Martin Jinek  Is a corresponding author
  1. University of Zurich, Switzerland
https://doi.org/10.7554/eLife.80332
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  1. Lena Maria Muckenfuss
  2. Anabel Carmen Migenda Herranz
  3. Franziska M Boneberg
  4. Marcello Clerici
  5. Martin Jinek
(2022)
Fip1 is a multivalent interaction scaffold for processing factors in human mRNA 3' end biogenesis
eLife 11:e80332.
https://doi.org/10.7554/eLife.80332
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Abstract

3' end formation of most eukaryotic mRNAs is dependent on the assembly of a ~1.5 megadalton multiprotein complex, that catalyzes the coupled reaction of pre-mRNA cleavage and polyadenylation. In mammals, the cleavage and polyadenylation specificity factor (CPSF) constitutes the core of the 3' end processing machinery onto which the remaining factors, including cleavage stimulation factor (CstF) and poly(A) polymerase (PAP), assemble. These interactions are mediated by Fip1, a CPSF subunit characterized by high degree of intrinsic disorder. Here, we report two crystal structures revealing the interactions of human Fip1 (hFip1) with CPSF30 and CstF77. We demonstrate that CPSF contains two copies of hFip1, each binding to the zinc finger (ZF) domains 4 and 5 of CPSF30. Using polyadenylation assays we show that the two hFip1 copies are functionally redundant in recruiting one copy of PAP, thereby increasing the processivity of RNA polyadenylation. We further show that the interaction between hFip1 and CstF77 is mediated via a short motif in the N-terminal 'acidic' region of hFip1. In turn, CstF77 competitively inhibits CPSF-dependent PAP recruitment and 3' polyadenylation. Taken together, these results provide a structural basis for the multivalent scaffolding and regulatory functions of hFip1 in 3' end processing.

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X-ray diffraction data (atomic coordinates and structure factors) have been submitted to the PDB and will be released upon publication.

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Lena Maria Muckenfuss

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

  2. Anabel Carmen Migenda Herranz

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

  3. Franziska M Boneberg

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

  4. Marcello Clerici

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

  5. 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

Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (NCCR RNA and Disease)

  • Martin Jinek

Boehringer Ingelheim Fonds

  • Lena Maria Muckenfuss

Howard Hughes Medical Institute (55008735)

  • Martin Jinek

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

Reviewing Editor

  1. Eric J Wagner, University of Rochester Medical Center, United States

Version history

  1. Received: May 17, 2022
  2. Preprint posted: May 31, 2022 (view preprint)
  3. Accepted: September 7, 2022
  4. Accepted Manuscript published: September 8, 2022 (version 1)
  5. Version of Record published: September 26, 2022 (version 2)

Copyright

© 2022, Muckenfuss 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. Lena Maria Muckenfuss
  2. Anabel Carmen Migenda Herranz
  3. Franziska M Boneberg
  4. Marcello Clerici
  5. Martin Jinek
(2022)
Fip1 is a multivalent interaction scaffold for processing factors in human mRNA 3' end biogenesis
eLife 11:e80332.
https://doi.org/10.7554/eLife.80332

Share this article

https://doi.org/10.7554/eLife.80332

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