Human RIF1 and protein phosphatase 1 stimulate DNA replication origin licensing but suppress origin activation

Shin-ichiro Hiraga, Tony Ly, Javier Garzon, Zuzana Horejsi, Yoshi-nobu Ohkubo, Akinori Endo, Chikashi Obuse, Simon J Boulton, Angus I Lamond, Anne D. Donaldson

Research output: Contribution to journalArticlepeer-review

Abstract

The human RIF1 protein controls DNA replication, but the molecular mechanism is largely unknown. Here, we demonstrate that human RIF1 negatively regulates DNA replication by forming a complex with protein phosphatase 1 (PP1) that limits phosphorylation-mediated activation of the MCM replicative helicase. We identify specific residues on four MCM helicase subunits that show hyperphosphorylation upon RIF1 depletion, with the regulatory N-terminal domain of MCM4 being particularly strongly affected. In addition to this role in limiting origin activation, we discover an unexpected new role for human RIF1-PP1 in mediating efficient origin licensing. Specifically, during the G1 phase of the cell cycle, RIF1-PP1 protects the origin-binding ORC1 protein from untimely phosphorylation and consequent degradation by the proteasome. Depletion of RIF1 or inhibition of PP1 destabilizes ORC1, thereby reducing origin licensing. Consistent with reduced origin licensing, RIF1-depleted cells exhibit increased spacing between active origins. Human RIF1 therefore acts as a PP1-targeting subunit that regulates DNA replication positively by stimulating the origin licensing step, and then negatively by counteracting replication origin activation.

Original languageEnglish
Pages (from-to)403-419
Number of pages17
JournalEMBO Reports
Volume18
Issue number3
Early online date11 Jan 2017
DOIs
Publication statusPublished - Mar 2017

Keywords

  • MCM
  • ORC1
  • origin licensing
  • PP1
  • RIF1
  • MEIER-GORLIN SYNDROME
  • EMBRYONIC STEM-CELLS
  • S-PHASE
  • RECOGNITION COMPLEX
  • DORMANT ORIGINS
  • END RESECTION
  • HUMAN INTERACTOME
  • MAMMALIAN-CELLS
  • TELOMERE LENGTH
  • TIMING PROGRAM

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