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EMBO Rep. 2017 Mar;18(3):403-419. doi: 10.15252/embr.201641983. Epub 2017 Jan 11.

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

Author information

1
Institute of Medical Sciences, School of Medicine, Medical Sciences & Nutrition, University of Aberdeen, Aberdeen, UK s.hiraga@abdn.ac.uk a.d.donaldson@abdn.ac.uk.
2
Centre for Gene Regulation & Expression, School of Life Sciences, University of Dundee, Dundee, UK.
3
Institute of Medical Sciences, School of Medicine, Medical Sciences & Nutrition, University of Aberdeen, Aberdeen, UK.
4
The Francis Crick Institute, Clare Hall Laboratories, South Mimms, UK.
5
Graduate School of Life Science, Hokkaido University, Sapporo Hokkaido, Japan.

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.

KEYWORDS:

MCM ; ORC1; PP1; RIF1; origin licensing

PMID:
28077461
PMCID:
PMC5331243
DOI:
10.15252/embr.201641983
[Indexed for MEDLINE]
Free PMC Article

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