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R-loops promote trinucleotide repeat deletion through DNA base excision repair enzymatic activities
- Source :
- J Biol Chem
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- Trinucleotide repeat (TNR) expansion and deletion are responsible for over 40 neurodegenerative diseases and associated with cancer. TNRs can undergo somatic instability that is mediated by DNA damage and repair and gene transcription. Recent studies have pointed toward a role for R-loops in causing TNR expansion and deletion, and it has been shown that base excision repair (BER) can result in CAG repeat deletion from R-loops in yeast. However, it remains unknown how BER in R-loops can mediate TNR instability. In this study, using biochemical approaches, we examined BER enzymatic activities and their influence on TNR R-loops. We found that AP endonuclease 1 incised an abasic site on the nontemplate strand of a TNR R-loop, creating a double-flap intermediate containing an RNA:DNA hybrid that subsequently inhibited polymerase β (pol β) synthesis of TNRs. This stimulated flap endonuclease 1 (FEN1) cleavage of TNRs engaged in an R-loop. Moreover, we showed that FEN1 also efficiently cleaved the RNA strand, facilitating pol β loop/hairpin bypass synthesis and the resolution of TNR R-loops through BER. Consequently, this resulted in fewer TNRs synthesized by pol β than those removed by FEN1, thereby leading to repeat deletion. Our results indicate that TNR R-loops preferentially lead to repeat deletion during BER by disrupting the balance between the addition and removal of TNRs. Our discoveries open a new avenue for the treatment and prevention of repeat expansion diseases and cancer.
- Subjects :
- 0301 basic medicine
DNA Repair
Flap Endonucleases
DNA repair
DNA polymerase
DNA damage
Flap structure-specific endonuclease 1
DNA and Chromosomes
Biochemistry
AP endonuclease
03 medical and health sciences
Trinucleotide Repeats
DNA-(Apurinic or Apyrimidinic Site) Lyase
Humans
AP site
Molecular Biology
DNA Polymerase beta
030102 biochemistry & molecular biology
biology
Chemistry
Cell Biology
Base excision repair
Cell biology
030104 developmental biology
biology.protein
R-Loop Structures
Trinucleotide repeat expansion
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 295
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi.dedup.....9819acdf7afc1065f7bf57ddfcdfec47