Protein phosphatase 1 acts as a RIF1 effector to suppress DSB resection prior to Shieldin action

Shin-Ya Isobe; Shin-ichiro Hiraga; Koji Nagao; Hiroyuki Sasanuma; Anne D. Donaldson; Chikashi Obuse

doi:10.1016/j.celrep.2021.109383

Protein phosphatase 1 acts as a RIF1 effector to suppress DSB resection prior to Shieldin action

Shin-Ya Isobe, Shin-ichiro Hiraga, Koji Nagao, Hiroyuki Sasanuma, Anne D. Donaldson, Chikashi Obuse^*

^*Corresponding author for this work

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

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Abstract

DNAdouble-strand breaks (DSBs) are repaired mainly by non-homologous end joining (NHEJ) or homologous recombination (HR). RIF1 negatively regulates resection through the effector Shieldin, which associates with a short 30 single-stranded DNA (ssDNA) overhang by the MRN (MRE11-RAD50-NBS1) complex, to prevent further resection and HR repair. In this study, we show that RIF1, but not Shieldin, inhibits the accumulation of CtIP at DSB sites immediately after damage, suggesting that RIF1 has another effector besides Shieldin. We find that protein phosphatase 1 (PP1), a known RIF1 effector in replication, localizes at damage sites dependent on RIF1, where it suppresses downstream CtIP accumulation and limits the resection by the MRN complex. PP1 therefore acts as a RIF1 effector distinct from Shieldin. Furthermore, PP1 deficiency in the context of Shieldin depletion elevates HR immediately after irradiation. We conclude that PP1 inhibits resection before the action of Shieldin to prevent precocious HR in the early phase of the damage response.

Original language	English
Article number	109383
Number of pages	18
Journal	Cell Reports
Volume	36
Issue number	2
DOIs	https://doi.org/10.1016/j.celrep.2021.109383
Publication status	Published - 13 Jul 2021

Bibliographical note

Funding Information:
We thank K. Murakami, H. Kimura, H. Kurumizaka, and J.M. Stark for materials. This work was supported by JSPS KAKENHI grant nos. JP19H03156 , JP18H04713 , JP18H05532 , and JP25116004 (to C.O.), JP17H06426 (to K.N.), JP18H04900 and JP19H04267 (to H.S.), the Mitsubishi Foundation (to H.S.), and by Cancer Research UK awards C1445/A19059 and DRCPGM/100013 (to S.H. and A.D.D. lab).

Publisher Copyright:
© 2021 The Author(s)

Keywords

RIF1
PP1
Resection
Shieldin
NHEJ
HR
Pathway Choice
CtiP
MRN
Olaparib

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Isobe_etal_CR_Protein_phosphatase_1_VOR
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Cite this

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title = "Protein phosphatase 1 acts as a RIF1 effector to suppress DSB resection prior to Shieldin action",

abstract = "DNAdouble-strand breaks (DSBs) are repaired mainly by non-homologous end joining (NHEJ) or homologous recombination (HR). RIF1 negatively regulates resection through the effector Shieldin, which associates with a short 30 single-stranded DNA (ssDNA) overhang by the MRN (MRE11-RAD50-NBS1) complex, to prevent further resection and HR repair. In this study, we show that RIF1, but not Shieldin, inhibits the accumulation of CtIP at DSB sites immediately after damage, suggesting that RIF1 has another effector besides Shieldin. We find that protein phosphatase 1 (PP1), a known RIF1 effector in replication, localizes at damage sites dependent on RIF1, where it suppresses downstream CtIP accumulation and limits the resection by the MRN complex. PP1 therefore acts as a RIF1 effector distinct from Shieldin. Furthermore, PP1 deficiency in the context of Shieldin depletion elevates HR immediately after irradiation. We conclude that PP1 inhibits resection before the action of Shieldin to prevent precocious HR in the early phase of the damage response.",

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author = "Shin-Ya Isobe and Shin-ichiro Hiraga and Koji Nagao and Hiroyuki Sasanuma and Donaldson, {Anne D.} and Chikashi Obuse",

note = "Funding Information: We thank K. Murakami, H. Kimura, H. Kurumizaka, and J.M. Stark for materials. This work was supported by JSPS KAKENHI grant nos. JP19H03156 , JP18H04713 , JP18H05532 , and JP25116004 (to C.O.), JP17H06426 (to K.N.), JP18H04900 and JP19H04267 (to H.S.), the Mitsubishi Foundation (to H.S.), and by Cancer Research UK awards C1445/A19059 and DRCPGM/100013 (to S.H. and A.D.D. lab). Publisher Copyright: {\textcopyright} 2021 The Author(s)",

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AU - Hiraga, Shin-ichiro

AU - Nagao, Koji

AU - Sasanuma, Hiroyuki

AU - Donaldson, Anne D.

AU - Obuse, Chikashi

N1 - Funding Information: We thank K. Murakami, H. Kimura, H. Kurumizaka, and J.M. Stark for materials. This work was supported by JSPS KAKENHI grant nos. JP19H03156 , JP18H04713 , JP18H05532 , and JP25116004 (to C.O.), JP17H06426 (to K.N.), JP18H04900 and JP19H04267 (to H.S.), the Mitsubishi Foundation (to H.S.), and by Cancer Research UK awards C1445/A19059 and DRCPGM/100013 (to S.H. and A.D.D. lab). Publisher Copyright: © 2021 The Author(s)

PY - 2021/7/13

Y1 - 2021/7/13

N2 - DNAdouble-strand breaks (DSBs) are repaired mainly by non-homologous end joining (NHEJ) or homologous recombination (HR). RIF1 negatively regulates resection through the effector Shieldin, which associates with a short 30 single-stranded DNA (ssDNA) overhang by the MRN (MRE11-RAD50-NBS1) complex, to prevent further resection and HR repair. In this study, we show that RIF1, but not Shieldin, inhibits the accumulation of CtIP at DSB sites immediately after damage, suggesting that RIF1 has another effector besides Shieldin. We find that protein phosphatase 1 (PP1), a known RIF1 effector in replication, localizes at damage sites dependent on RIF1, where it suppresses downstream CtIP accumulation and limits the resection by the MRN complex. PP1 therefore acts as a RIF1 effector distinct from Shieldin. Furthermore, PP1 deficiency in the context of Shieldin depletion elevates HR immediately after irradiation. We conclude that PP1 inhibits resection before the action of Shieldin to prevent precocious HR in the early phase of the damage response.

AB - DNAdouble-strand breaks (DSBs) are repaired mainly by non-homologous end joining (NHEJ) or homologous recombination (HR). RIF1 negatively regulates resection through the effector Shieldin, which associates with a short 30 single-stranded DNA (ssDNA) overhang by the MRN (MRE11-RAD50-NBS1) complex, to prevent further resection and HR repair. In this study, we show that RIF1, but not Shieldin, inhibits the accumulation of CtIP at DSB sites immediately after damage, suggesting that RIF1 has another effector besides Shieldin. We find that protein phosphatase 1 (PP1), a known RIF1 effector in replication, localizes at damage sites dependent on RIF1, where it suppresses downstream CtIP accumulation and limits the resection by the MRN complex. PP1 therefore acts as a RIF1 effector distinct from Shieldin. Furthermore, PP1 deficiency in the context of Shieldin depletion elevates HR immediately after irradiation. We conclude that PP1 inhibits resection before the action of Shieldin to prevent precocious HR in the early phase of the damage response.

KW - RIF1

KW - PP1

KW - Resection

KW - Shieldin

KW - NHEJ

KW - HR

KW - Pathway Choice

KW - CtiP

KW - MRN

KW - Olaparib

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DO - 10.1016/j.celrep.2021.109383

M3 - Article

SN - 2211-1247

VL - 36

JO - Cell Reports

JF - Cell Reports

IS - 2

M1 - 109383

ER -

Protein phosphatase 1 acts as a RIF1 effector to suppress DSB resection prior to Shieldin action

Abstract

Bibliographical note

Keywords

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