BRCA2 associates with MCM10 to suppress PRIMPOL-mediated repriming and single-stranded gap formation after DNA damage

Zhihua Kang; Pan Fu; Allen L. Alcivar; Haiqing Fu; Christophe Redon; Tzeh Keong Foo; Yamei Zuo; Caiyong Ye; Ryan Baxley; Advaitha Madireddy; Remi Buisson; Anja Katrin Bielinsky; Lee Zou; Zhiyuan Shen; Mirit I. Aladjem; Bing Xia

doi:10.1038/s41467-021-26227-6

BRCA2 associates with MCM10 to suppress PRIMPOL-mediated repriming and single-stranded gap formation after DNA damage

Zhihua Kang, Pan Fu, Allen L. Alcivar, Haiqing Fu, Christophe Redon, Tzeh Keong Foo, Yamei Zuo, Caiyong Ye, Ryan Baxley, Advaitha Madireddy, Remi Buisson, Anja Katrin Bielinsky, Lee Zou, Zhiyuan Shen, Mirit I. Aladjem, Bing Xia

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

54 Scopus citations

Abstract

The BRCA2 tumor suppressor protects genome integrity by promoting homologous recombination-based repair of DNA breaks, stability of stalled DNA replication forks and DNA damage-induced cell cycle checkpoints. BRCA2 deficient cells display the radio-resistant DNA synthesis (RDS) phenotype, however the mechanism has remained elusive. Here we show that cells without BRCA2 are unable to sufficiently restrain DNA replication fork progression after DNA damage, and the underrestrained fork progression is due primarily to Primase-Polymerase (PRIMPOL)-mediated repriming of DNA synthesis downstream of lesions, leaving behind single-stranded DNA gaps. Moreover, we find that BRCA2 associates with the essential DNA replication factor MCM10 and this association suppresses PRIMPOL-mediated repriming and ssDNA gap formation, while having no impact on the stability of stalled replication forks. Our findings establish an important function for BRCA2, provide insights into replication fork control during the DNA damage response, and may have implications in tumor suppression and therapy response.

Original language	English (US)
Article number	5966
Journal	Nature communications
Volume	12
Issue number	1
DOIs	https://doi.org/10.1038/s41467-021-26227-6
State	Published - Dec 1 2021

Bibliographical note

Funding Information:
We thank Dr. Juan Mendez (Spanish National Cancer Research Centre) for provision of the PRIMPOL antibody and cDNA constructs, Dr. Maria Jasin (Memorial Sloan Kettering Cancer Center) for providing VC8 and VC8 + BRCA2 cells, and Dr. Inder Verma (Salk Institute) for providing the pMG-BRCA2-GFP plasmid. This work was supported by the US National Cancer Institute (R01CA138804, R01CA262227, and P01CA250957 to Z.S. and B.X.), the Robert Wood Johnson Foundation (through a grant to Rutgers Cancer Institute of New Jersey). Flow cytometry data were generated using the Rutgers Cancer Institute of New Jersey Flow Cytometry and Cell Sorting Shared Resource, which is supported, in part, by NCI-CCSG P30CA072720-5921. Z.K. and T.K.F were supported by postdoctoral fellowships from the New Jersey Commission on Cancer Research (NJCCR).

Publisher Copyright:
© 2021, The Author(s).

Keywords

BRCA2 Protein/antagonists & inhibitors
Cell Line, Tumor
Cell Survival
DNA Damage
DNA Helicases/antagonists & inhibitors
DNA Primase/antagonists & inhibitors
DNA Replication
DNA, Neoplasm/genetics
DNA, Single-Stranded/genetics
DNA-Binding Proteins/antagonists & inhibitors
DNA-Directed DNA Polymerase/genetics
Gene Expression Regulation, Neoplastic
Genomic Instability
HEK293 Cells
HeLa Cells
Humans
Minichromosome Maintenance Proteins/antagonists & inhibitors
Multifunctional Enzymes/antagonists & inhibitors
Osteoblasts/metabolism
RNA, Small Interfering/genetics
Recombinational DNA Repair
Signal Transduction
Transcription Factors/antagonists & inhibitors

PubMed: MeSH publication types

Research Support, Non-U.S. Gov't
Journal Article
Research Support, N.I.H., Extramural

UN SDGs

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

Publisher link

10.1038/s41467-021-26227-6

Find It

Find It at U of M Libraries

Cite this

Kang, Z., Fu, P., Alcivar, A. L., Fu, H., Redon, C., Foo, T. K., Zuo, Y., Ye, C., Baxley, R., Madireddy, A., Buisson, R., Bielinsky, A. K., Zou, L., Shen, Z., Aladjem, M. I., & Xia, B. (2021). BRCA2 associates with MCM10 to suppress PRIMPOL-mediated repriming and single-stranded gap formation after DNA damage. Nature communications, 12(1), Article 5966. https://doi.org/10.1038/s41467-021-26227-6

Kang, Z, Fu, P, Alcivar, AL, Fu, H, Redon, C, Foo, TK, Zuo, Y, Ye, C, Baxley, R, Madireddy, A, Buisson, R, Bielinsky, AK, Zou, L, Shen, Z, Aladjem, MI & Xia, B 2021, 'BRCA2 associates with MCM10 to suppress PRIMPOL-mediated repriming and single-stranded gap formation after DNA damage', Nature communications, vol. 12, no. 1, 5966. https://doi.org/10.1038/s41467-021-26227-6

@article{50f2a72ca3ff401aaad321d0a5f7e361,

title = "BRCA2 associates with MCM10 to suppress PRIMPOL-mediated repriming and single-stranded gap formation after DNA damage",

abstract = "The BRCA2 tumor suppressor protects genome integrity by promoting homologous recombination-based repair of DNA breaks, stability of stalled DNA replication forks and DNA damage-induced cell cycle checkpoints. BRCA2 deficient cells display the radio-resistant DNA synthesis (RDS) phenotype, however the mechanism has remained elusive. Here we show that cells without BRCA2 are unable to sufficiently restrain DNA replication fork progression after DNA damage, and the underrestrained fork progression is due primarily to Primase-Polymerase (PRIMPOL)-mediated repriming of DNA synthesis downstream of lesions, leaving behind single-stranded DNA gaps. Moreover, we find that BRCA2 associates with the essential DNA replication factor MCM10 and this association suppresses PRIMPOL-mediated repriming and ssDNA gap formation, while having no impact on the stability of stalled replication forks. Our findings establish an important function for BRCA2, provide insights into replication fork control during the DNA damage response, and may have implications in tumor suppression and therapy response.",

keywords = "BRCA2 Protein/antagonists & inhibitors, Cell Line, Tumor, Cell Survival, DNA Damage, DNA Helicases/antagonists & inhibitors, DNA Primase/antagonists & inhibitors, DNA Replication, DNA, Neoplasm/genetics, DNA, Single-Stranded/genetics, DNA-Binding Proteins/antagonists & inhibitors, DNA-Directed DNA Polymerase/genetics, Gene Expression Regulation, Neoplastic, Genomic Instability, HEK293 Cells, HeLa Cells, Humans, Minichromosome Maintenance Proteins/antagonists & inhibitors, Multifunctional Enzymes/antagonists & inhibitors, Osteoblasts/metabolism, RNA, Small Interfering/genetics, Recombinational DNA Repair, Signal Transduction, Transcription Factors/antagonists & inhibitors",

author = "Zhihua Kang and Pan Fu and Alcivar, {Allen L.} and Haiqing Fu and Christophe Redon and Foo, {Tzeh Keong} and Yamei Zuo and Caiyong Ye and Ryan Baxley and Advaitha Madireddy and Remi Buisson and Bielinsky, {Anja Katrin} and Lee Zou and Zhiyuan Shen and Aladjem, {Mirit I.} and Bing Xia",

note = "Funding Information: We thank Dr. Juan Mendez (Spanish National Cancer Research Centre) for provision of the PRIMPOL antibody and cDNA constructs, Dr. Maria Jasin (Memorial Sloan Kettering Cancer Center) for providing VC8 and VC8 + BRCA2 cells, and Dr. Inder Verma (Salk Institute) for providing the pMG-BRCA2-GFP plasmid. This work was supported by the US National Cancer Institute (R01CA138804, R01CA262227, and P01CA250957 to Z.S. and B.X.), the Robert Wood Johnson Foundation (through a grant to Rutgers Cancer Institute of New Jersey). Flow cytometry data were generated using the Rutgers Cancer Institute of New Jersey Flow Cytometry and Cell Sorting Shared Resource, which is supported, in part, by NCI-CCSG P30CA072720-5921. Z.K. and T.K.F were supported by postdoctoral fellowships from the New Jersey Commission on Cancer Research (NJCCR). Publisher Copyright: {\textcopyright} 2021, The Author(s).",

year = "2021",

month = dec,

day = "1",

doi = "10.1038/s41467-021-26227-6",

language = "English (US)",

volume = "12",

journal = "Nature communications",

issn = "2041-1723",

publisher = "Nature Research",

number = "1",

}

TY - JOUR

T1 - BRCA2 associates with MCM10 to suppress PRIMPOL-mediated repriming and single-stranded gap formation after DNA damage

AU - Kang, Zhihua

AU - Fu, Pan

AU - Alcivar, Allen L.

AU - Fu, Haiqing

AU - Redon, Christophe

AU - Foo, Tzeh Keong

AU - Zuo, Yamei

AU - Ye, Caiyong

AU - Baxley, Ryan

AU - Madireddy, Advaitha

AU - Buisson, Remi

AU - Bielinsky, Anja Katrin

AU - Zou, Lee

AU - Shen, Zhiyuan

AU - Aladjem, Mirit I.

AU - Xia, Bing

N1 - Funding Information: We thank Dr. Juan Mendez (Spanish National Cancer Research Centre) for provision of the PRIMPOL antibody and cDNA constructs, Dr. Maria Jasin (Memorial Sloan Kettering Cancer Center) for providing VC8 and VC8 + BRCA2 cells, and Dr. Inder Verma (Salk Institute) for providing the pMG-BRCA2-GFP plasmid. This work was supported by the US National Cancer Institute (R01CA138804, R01CA262227, and P01CA250957 to Z.S. and B.X.), the Robert Wood Johnson Foundation (through a grant to Rutgers Cancer Institute of New Jersey). Flow cytometry data were generated using the Rutgers Cancer Institute of New Jersey Flow Cytometry and Cell Sorting Shared Resource, which is supported, in part, by NCI-CCSG P30CA072720-5921. Z.K. and T.K.F were supported by postdoctoral fellowships from the New Jersey Commission on Cancer Research (NJCCR). Publisher Copyright: © 2021, The Author(s).

PY - 2021/12/1

Y1 - 2021/12/1

N2 - The BRCA2 tumor suppressor protects genome integrity by promoting homologous recombination-based repair of DNA breaks, stability of stalled DNA replication forks and DNA damage-induced cell cycle checkpoints. BRCA2 deficient cells display the radio-resistant DNA synthesis (RDS) phenotype, however the mechanism has remained elusive. Here we show that cells without BRCA2 are unable to sufficiently restrain DNA replication fork progression after DNA damage, and the underrestrained fork progression is due primarily to Primase-Polymerase (PRIMPOL)-mediated repriming of DNA synthesis downstream of lesions, leaving behind single-stranded DNA gaps. Moreover, we find that BRCA2 associates with the essential DNA replication factor MCM10 and this association suppresses PRIMPOL-mediated repriming and ssDNA gap formation, while having no impact on the stability of stalled replication forks. Our findings establish an important function for BRCA2, provide insights into replication fork control during the DNA damage response, and may have implications in tumor suppression and therapy response.

AB - The BRCA2 tumor suppressor protects genome integrity by promoting homologous recombination-based repair of DNA breaks, stability of stalled DNA replication forks and DNA damage-induced cell cycle checkpoints. BRCA2 deficient cells display the radio-resistant DNA synthesis (RDS) phenotype, however the mechanism has remained elusive. Here we show that cells without BRCA2 are unable to sufficiently restrain DNA replication fork progression after DNA damage, and the underrestrained fork progression is due primarily to Primase-Polymerase (PRIMPOL)-mediated repriming of DNA synthesis downstream of lesions, leaving behind single-stranded DNA gaps. Moreover, we find that BRCA2 associates with the essential DNA replication factor MCM10 and this association suppresses PRIMPOL-mediated repriming and ssDNA gap formation, while having no impact on the stability of stalled replication forks. Our findings establish an important function for BRCA2, provide insights into replication fork control during the DNA damage response, and may have implications in tumor suppression and therapy response.

KW - BRCA2 Protein/antagonists & inhibitors

KW - Cell Line, Tumor

KW - Cell Survival

KW - DNA Damage

KW - DNA Helicases/antagonists & inhibitors

KW - DNA Primase/antagonists & inhibitors

KW - DNA Replication

KW - DNA, Neoplasm/genetics

KW - DNA, Single-Stranded/genetics

KW - DNA-Binding Proteins/antagonists & inhibitors

KW - DNA-Directed DNA Polymerase/genetics

KW - Gene Expression Regulation, Neoplastic

KW - Genomic Instability

KW - HEK293 Cells

KW - HeLa Cells

KW - Humans

KW - Minichromosome Maintenance Proteins/antagonists & inhibitors

KW - Multifunctional Enzymes/antagonists & inhibitors

KW - Osteoblasts/metabolism

KW - RNA, Small Interfering/genetics

KW - Recombinational DNA Repair

KW - Signal Transduction

KW - Transcription Factors/antagonists & inhibitors

UR - http://www.scopus.com/inward/record.url?scp=85117410055&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85117410055&partnerID=8YFLogxK

U2 - 10.1038/s41467-021-26227-6

DO - 10.1038/s41467-021-26227-6

M3 - Article

C2 - 34645815

AN - SCOPUS:85117410055

SN - 2041-1723

VL - 12

JO - Nature communications

JF - Nature communications

IS - 1

M1 - 5966

ER -

BRCA2 associates with MCM10 to suppress PRIMPOL-mediated repriming and single-stranded gap formation after DNA damage

Abstract

Bibliographical note

Keywords

PubMed: MeSH publication types

UN SDGs

Publisher link

Find It

Other files and links

Fingerprint

Cite this