The RIF1-Long splice variant promotes G1 phase 53BP1 nuclear bodies to protect against replication stress

Lotte P Watts; Toyoaki Natsume; Yuichiro Saito; Javier Garzon; Qianqian Dong; Lora Boteva; Nick Gilbert; Masato T Kanemaki; Shin-Ichiro Hiraga; Anne D Donaldson

doi:10.7554/eLife.58020

The RIF1-Long splice variant promotes G1 phase 53BP1 nuclear bodies to protect against replication stress

Lotte P Watts, Toyoaki Natsume, Yuichiro Saito, Javier Garzon, Qianqian Dong, Lora Boteva, Nick Gilbert, Masato T Kanemaki, Shin-Ichiro Hiraga, Anne D Donaldson

Research output: Contribution to journal › Article › peer-review

11 Citations (Scopus)

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Abstract

Human cells lacking RIF1 are highly sensitive to replication inhibitors, but the reasons for this sensitivity have been enigmatic. Here we show that RIF1 must be present both during replication stress and in the ensuing recovery period to promote cell survival. Of two isoforms produced by alternative splicing, we find that RIF1-Long alone can protect cells against replication inhibition, but RIF1-Short is incapable of mediating protection. Consistent with this isoform-specific role, RIF1-Long is required to promote the formation of the 53BP1 nuclear bodies that protect unrepaired damage sites in the G1 phase following replication stress. Overall, our observations show that RIF1 is needed at several cell cycle stages after replication insult, with the RIF1-Long isoform playing a specific role during the ensuing G1 phase in damage site protection.

Original language	English
Article number	e58020
Number of pages	26
Journal	eLife
Volume	9
Early online date	3 Nov 2020
DOIs	https://doi.org/10.7554/eLife.58020
Publication status	Published - 3 Nov 2020

Bibliographical note

Acknowledgements
Thanks to members of the Aberdeen Chromosome Biology Group for helpful comments, and Ronan Broderick and Wojciech Niedzwiedz for advice on mitotic bridge analysis. We thank Raif Yuecel and his team at the Iain Fraser Cytometry Centre for assistance, and Kevin Mackenzie and his team at the Microscopy and Histology Core Facility. Work was supported by Cancer Research UK Studentship Award C1445/A20596 and CRUK Programme Award C1445/A19059; by JSPS KAKENHI Grants Numbers 17K15068, 18H02170 and 18H04719; by research grants from the Daiichi Sankyo’s Foundation of Life Science and the Takeda Science Foundation; and by the UK Medical Research Council (MC_UU_00007/13). Collaboration was supported by a 2017 JSPS Summer Programme Fellowship.
Funding
Cancer Research UK (C1445/A20596)
Anne D Donaldson
Cancer Research UK (C1445/A19059)
Anne D Donaldson
Japan Society for the Promotion of Science (17K15068)
Masato T Kanemaki
Japan Society for the Promotion of Science (18H02170)
Masato T Kanemaki
Japan Society for the Promotion of Science (18H04719)
Masato T Kanemaki
Medical Research Council (MC_UU_00007/13)
Nick Gilbert

Keywords

RIF1
DNA replication
DNA replication stress
Splicing
replication stress
cell biology
chromosomes
splicing
human
gene expression

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.7554/eLife.58020Licence: CC BY

Watts_et_al_RIF1_long-splice_elife_vor
Copyright Watts et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited. https://creativecommons.org/licenses/by/4.0/
Final published version, 2.34 MBLicence: CC BY

Cite this

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title = "The RIF1-Long splice variant promotes G1 phase 53BP1 nuclear bodies to protect against replication stress",

abstract = "Human cells lacking RIF1 are highly sensitive to replication inhibitors, but the reasons for this sensitivity have been enigmatic. Here we show that RIF1 must be present both during replication stress and in the ensuing recovery period to promote cell survival. Of two isoforms produced by alternative splicing, we find that RIF1-Long alone can protect cells against replication inhibition, but RIF1-Short is incapable of mediating protection. Consistent with this isoform-specific role, RIF1-Long is required to promote the formation of the 53BP1 nuclear bodies that protect unrepaired damage sites in the G1 phase following replication stress. Overall, our observations show that RIF1 is needed at several cell cycle stages after replication insult, with the RIF1-Long isoform playing a specific role during the ensuing G1 phase in damage site protection. ",

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note = "Acknowledgements Thanks to members of the Aberdeen Chromosome Biology Group for helpful comments, and Ronan Broderick and Wojciech Niedzwiedz for advice on mitotic bridge analysis. We thank Raif Yuecel and his team at the Iain Fraser Cytometry Centre for assistance, and Kevin Mackenzie and his team at the Microscopy and Histology Core Facility. Work was supported by Cancer Research UK Studentship Award C1445/A20596 and CRUK Programme Award C1445/A19059; by JSPS KAKENHI Grants Numbers 17K15068, 18H02170 and 18H04719; by research grants from the Daiichi Sankyo{\textquoteright}s Foundation of Life Science and the Takeda Science Foundation; and by the UK Medical Research Council (MC_UU_00007/13). Collaboration was supported by a 2017 JSPS Summer Programme Fellowship. Funding Cancer Research UK (C1445/A20596) Anne D Donaldson Cancer Research UK (C1445/A19059) Anne D Donaldson Japan Society for the Promotion of Science (17K15068) Masato T Kanemaki Japan Society for the Promotion of Science (18H02170) Masato T Kanemaki Japan Society for the Promotion of Science (18H04719) Masato T Kanemaki Medical Research Council (MC_UU_00007/13) Nick Gilbert",

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AU - Watts, Lotte P

AU - Natsume, Toyoaki

AU - Saito, Yuichiro

AU - Garzon, Javier

AU - Dong, Qianqian

AU - Boteva, Lora

AU - Gilbert, Nick

AU - Kanemaki, Masato T

AU - Hiraga, Shin-Ichiro

AU - Donaldson, Anne D

N1 - Acknowledgements Thanks to members of the Aberdeen Chromosome Biology Group for helpful comments, and Ronan Broderick and Wojciech Niedzwiedz for advice on mitotic bridge analysis. We thank Raif Yuecel and his team at the Iain Fraser Cytometry Centre for assistance, and Kevin Mackenzie and his team at the Microscopy and Histology Core Facility. Work was supported by Cancer Research UK Studentship Award C1445/A20596 and CRUK Programme Award C1445/A19059; by JSPS KAKENHI Grants Numbers 17K15068, 18H02170 and 18H04719; by research grants from the Daiichi Sankyo’s Foundation of Life Science and the Takeda Science Foundation; and by the UK Medical Research Council (MC_UU_00007/13). Collaboration was supported by a 2017 JSPS Summer Programme Fellowship. Funding Cancer Research UK (C1445/A20596) Anne D Donaldson Cancer Research UK (C1445/A19059) Anne D Donaldson Japan Society for the Promotion of Science (17K15068) Masato T Kanemaki Japan Society for the Promotion of Science (18H02170) Masato T Kanemaki Japan Society for the Promotion of Science (18H04719) Masato T Kanemaki Medical Research Council (MC_UU_00007/13) Nick Gilbert

PY - 2020/11/3

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N2 - Human cells lacking RIF1 are highly sensitive to replication inhibitors, but the reasons for this sensitivity have been enigmatic. Here we show that RIF1 must be present both during replication stress and in the ensuing recovery period to promote cell survival. Of two isoforms produced by alternative splicing, we find that RIF1-Long alone can protect cells against replication inhibition, but RIF1-Short is incapable of mediating protection. Consistent with this isoform-specific role, RIF1-Long is required to promote the formation of the 53BP1 nuclear bodies that protect unrepaired damage sites in the G1 phase following replication stress. Overall, our observations show that RIF1 is needed at several cell cycle stages after replication insult, with the RIF1-Long isoform playing a specific role during the ensuing G1 phase in damage site protection.

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KW - RIF1

KW - DNA replication

KW - DNA replication stress

KW - Splicing

KW - replication stress

KW - cell biology

KW - chromosomes

KW - splicing

KW - human

KW - gene expression

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U2 - 10.7554/eLife.58020

DO - 10.7554/eLife.58020

M3 - Article

C2 - 33141022

VL - 9

JO - eLife

JF - eLife

M1 - e58020

ER -

The RIF1-Long splice variant promotes G1 phase 53BP1 nuclear bodies to protect against replication stress

Abstract

Bibliographical note

Keywords

UN SDGs

Access to Document

Other files and links

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Iain Fraser Cytometry Centre

Microscopy and Histology

Cite this

The RIF1-Long splice variant promotes G1 phase 53BP1 nuclear bodies to protect against replication stress

Abstract

Bibliographical note

Keywords

UN SDGs

Access to Document

Other files and links

Fingerprint

Equipment

Iain Fraser Cytometry Centre

Microscopy and Histology

Cite this