Mouse Rif1 is a regulatory subunit of protein phosphatase 1 (PP1)

Scientific Reports
Rasa SukackaiteSara B Buonomo

Abstract

Rif1 is a conserved protein that plays essential roles in orchestrating DNA replication timing, controlling nuclear architecture, telomere length and DNA repair. However, the relationship between these different roles, as well as the molecular basis of Rif1 function is still unclear. The association of Rif1 with insoluble nuclear lamina has thus far hampered exhaustive characterization of the associated protein complexes. We devised a protocol that overcomes this problem, and were thus able to discover a number of novel Rif1 interactors, involved in chromatin metabolism and phosphorylation. Among them, we focus here on PP1. Data from different systems have suggested that Rif1-PP1 interaction is conserved and has important biological roles. Using mutagenesis, NMR, isothermal calorimetry and surface plasmon resonance we demonstrate that Rif1 is a high-affinity PP1 adaptor, able to out-compete the well-established PP1-inhibitor I2 in vitro. Our conclusions have important implications for understanding Rif1 diverse roles and the relationship between the biological processes controlled by Rif1.

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Citations

Mar 9, 2019·Genes·Dominik Boos, Pedro Ferreira
Aug 15, 2018·EMBO Reports·Shin-Ichiro HiragaAnne D Donaldson
Sep 4, 2019·Nature Reviews. Molecular Cell Biology·Claire MarchalDavid M Gilbert
May 11, 2018·PLoS Biology·Charles A Seller, Patrick H O'Farrell
Jul 25, 2019·Nature Communications·Chirantani MukherjeeArnab Ray Chaudhuri
Jul 16, 2019·Cellular and Molecular Life Sciences : CMLS·Sandra Segura-Bayona, Travis H Stracker
Apr 2, 2020·Nature Structural & Molecular Biology·Aaron Mendez-BermudezEric Gilson
Jul 28, 2020·Trends in Genetics : TIG·Patroula NathanailidouZoi Lygerou
Nov 11, 2020·Journal of Biochemistry·Sana AlaviHisao Masai
May 20, 2021·Nature Communications·Stefano GnanSara C B Buonomo
Aug 3, 2020·Trends in Genetics : TIG·Qiliang Ding, Amnon Koren
Sep 2, 2021·Nucleic Acids Research·Diletta CiardoKathrin Marheineke
Nov 18, 2021·The EMBO Journal·Elin EnervaldSara B C Buonomo

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Methods Mentioned

BETA
affinity
immunoprecipitation
surface plasmon resonance
size exclusion chromatography
gel-filtration
NMR
isothermal titration calorimetry
chip
dissection
electrophoresis

Software Mentioned

Panther
ESPRIT
Perseus
MaxQuant LFQ
BIAevaluation
BioMart
MaxQuant

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