Ligase 3–mediated end-joining maintains genome stability of human embryonic stem cells

Aneta Kohutova, Jan Raška, Miriama Kruta, Monika Seneklova, Tomas Barta, Petr Fojtik, Tereza Jurakova, Christi A. Walter, Ales Hampl, Petr Dvorak, Vladimir Rotrekl

Research output: Contribution to journalArticle

Abstract

Maintenance of human embryonic stem cells (hESCs) with stable genome is important for their future use in cell replacement therapy and disease modeling. Our understanding of the mechanisms maintaining genomic stability of hESC and our ability to modulate them is essential in preventing unwanted mutation accumulation during their in vitro cultivation. In this study, we show the DNA damage response mechanism in hESCs is composed of known, yet unlikely components. Clustered oxidative base damage is converted into DNA double-strand breaks (DSBs) by base excision repair (BER) and then quickly repaired by ligase (Lig)3-mediated end-joining (EJ). If there is further induction of clustered oxidative base damage by irradiation, then BER-mediated DSBs become essential in triggering the checkpoint response in hESCs. hESCs limit the mutagenic potential of Lig3–mediated EJ by DNA break end protection involving p53 binding protein 1 (53BP1), which results in fast and error-free microhomology-mediated repair and a low mutant frequency in hESCs. DSBs in hESCs are also repaired via homologous recombination (HR); however, DSB overload, together with massive end protection by 53BP1, triggers competition between error-free HR and mutagenic nonhomologous EJ.—Kohutova, A., Raška, J., Kruta, M., Seneklova, M., Barta, T., Fojtik, P., Jurakova, T., Walter, C. A., Hampl, A., Dvorak, P., Rotrekl, V. Ligase 3–mediated end-joining maintains genome stability of human embryonic stem cells. FASEB J. 33, 6778–6788 (2019). www.fasebj.org.

Original languageEnglish (US)
Pages (from-to)6778-6788
Number of pages11
JournalFASEB Journal
Volume33
Issue number6
DOIs
StatePublished - Jun 1 2019

Keywords

  • 53BP1
  • PARP1
  • alternative DNA end-joining
  • base excision repair
  • pluripotent stem cells

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'Ligase 3–mediated end-joining maintains genome stability of human embryonic stem cells'. Together they form a unique fingerprint.

  • Cite this

    Kohutova, A., Raška, J., Kruta, M., Seneklova, M., Barta, T., Fojtik, P., Jurakova, T., Walter, C. A., Hampl, A., Dvorak, P., & Rotrekl, V. (2019). Ligase 3–mediated end-joining maintains genome stability of human embryonic stem cells. FASEB Journal, 33(6), 6778-6788. https://doi.org/10.1096/fj.201801877RR