TY - JOUR
T1 - Ligase 3–mediated end-joining maintains genome stability of human embryonic stem cells
AU - Kohutova, Aneta
AU - Raška, Jan
AU - Kruta, Miriama
AU - Seneklova, Monika
AU - Barta, Tomas
AU - Fojtik, Petr
AU - Jurakova, Tereza
AU - Walter, Christi A.
AU - Hampl, Ales
AU - Dvorak, Petr
AU - Rotrekl, Vladimir
N1 - Funding Information:
This work was supported by the Grant Agency of the Czech Republic (GACR; Grant P302/12/G157), the National Program of Sustainability II (MEYS CR; LQ1605), and the European Research Center for Life Science, Advanced Materials and Nanotechnology (CEITEC 2020; LQ1601). This work was also supported by the Rector's Program for Student Projects from Masaryk University (MUNI/C/0967/2013). The authors declare no conflicts of interest.
Publisher Copyright:
© FASEB
PY - 2019/6/1
Y1 - 2019/6/1
N2 - 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.
AB - 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.
KW - 53BP1
KW - PARP1
KW - alternative DNA end-joining
KW - base excision repair
KW - pluripotent stem cells
UR - http://www.scopus.com/inward/record.url?scp=85067276713&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85067276713&partnerID=8YFLogxK
U2 - 10.1096/fj.201801877RR
DO - 10.1096/fj.201801877RR
M3 - Article
C2 - 30807703
AN - SCOPUS:85067276713
SN - 0892-6638
VL - 33
SP - 6778
EP - 6788
JO - FASEB Journal
JF - FASEB Journal
IS - 6
ER -