TDP1 is required for efficient non-homologous end joining in human cells

DNA Repair
Jing LiLeslyn A Hanakahi

Abstract

Tyrosyl-DNA phosphodiesterase 1 (TDP1) can remove a wide variety of 3' and 5' terminal DNA adducts. Genetic studies in yeast identified TDP1 as a regulator of non-homologous end joining (NHEJ) fidelity in the repair of double-strand breaks (DSBs) lacking terminal adducts. In this communication, we show that TDP1 plays an important role in joining cohesive DSBs in human cells. To investigate the role of TDP1 in NHEJ in live human cells we used CRISPR/cas9 to produce TDP1-knockout (TDP1-KO) HEK-293 cells. As expected, human TDP1-KO cells were highly sensitive to topoisomerase poisons and ionizing radiation. Using a chromosomally-integrated NHEJ reporter substrate to compare end joining between wild type and TDP1-KO cells, we found that TDP1-KO cells have a 5-fold reduced ability to repair I-SceI-generated DSBs. Extracts prepared from TDP1-KO cells had reduced NHEJ activity in vitro, as compared to extracts from wild type cells. Analysis of end-joining junctions showed that TDP1 deficiency reduced end-joining fidelity, with a significant increase in insertion events, similar to previous observations in yeast. It has been reported that phosphorylation of TDP1 serine 81 (TDP1-S81) by ATM and DNA-PK stabilizes TDP1 and recruits TDP1 ...Continue Reading

Citations

Dec 8, 2017·Nucleic Acids Research·Ajinkya S Kawale, Lawrence F Povirk
Aug 6, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Irina V Il'inaOlga I Lavrik
Aug 23, 2019·DNA Repair·Karin C NitissLeslyn A Hanakahi
May 31, 2019·Emerging Topics in Life Sciences·Marianna RomitoAlessia Cavazza
Feb 7, 2021·Biochemical and Biophysical Research Communications·Ben YangTao Li
Jul 25, 2021·Sensors·Ann-Katrine JakobsenMagnus Stougaard

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