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 journalArticlepeer-review

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 languageEnglish (US)
Article number5966
JournalNature communications
Volume12
Issue number1
DOIs
StatePublished - Dec 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

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