Related Experiment Video

Updated: Oct 18, 2025

Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro
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Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro

Published on: May 3, 2014

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Dynamics in Fip1 regulate eukaryotic mRNA 3' end processing

Ananthanarayanan Kumar1, Conny W H Yu1, Juan B Rodríguez-Molina1

  • 1MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.

Genes & Development
|October 1, 2021
PubMed

Insights

The study reveals how yeast Fip1 anchors poly(A) polymerase Pap1 to the cleavage and polyadenylation factor (CPF) complex. Fip1

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • The cleavage and polyadenylation factor (CPF) complex is crucial for mRNA 3' end processing in eukaryotes.
  • CPF comprises multiple subunits, some of which possess intrinsically disordered regions (IDRs).
  • Intrinsically disordered regions can exhibit flexibility or adopt ordered structures upon binding partners.

Purpose of the Study:

  • To investigate the role of intrinsically disordered regions in CPF function.
  • To elucidate the interaction between Fip1, Pap1, and Yth1 within the CPF complex.
  • To determine the dynamics of Fip1 within the CPF complex during mRNA processing.

Main Methods:

  • Reconstitution of a recombinant 850-kDa CPF complex.
  • Selective labeling of Fip1 for incorporation into the recombinant CPF.
  • Nuclear magnetic resonance (NMR) spectroscopy to study Fip1 dynamics within CPF.

Main Results:

  • Yeast Fip1 anchors the poly(A) polymerase Pap1 to CPF through an interaction with Yth1's zinc finger 4.
  • A Fip1 intrinsically disordered region connecting Yth1- and Pap1-binding sites remains highly dynamic within the CPF complex.
  • The dynamics of Fip1 are essential for coordinating cleavage and polyadenylation.

Conclusions:

  • Fip1's dynamic intrinsically disordered region is key to its function in CPF.
  • The flexibility of Fip1 facilitates the coordination of mRNA cleavage and polyadenylation.
  • Understanding Fip1 dynamics provides insights into the regulation of mRNA 3' end processing.
Keywords:
CPFCPSFRNA-binding proteindynamicsmRNA processingpolyadenylation

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