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One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
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Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
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Flap Endonuclease 1 Endonucleolytically Processes RNA to Resolve R-Loops through DNA Base Excision Repair.

Eduardo E Laverde1, Aris A Polyzos2, Pawlos P Tsegay1

  • 1Biochemistry Ph.D. Program, Florida International University, Miami, FL 33199, USA.

Genes
|January 21, 2023
PubMed
Summary

Flap endonuclease 1 (FEN1) resolves R-loops by cleaving RNA strands during DNA repair. This enzyme

Keywords:
R-loopbase excision repair (BER)flap endonuclease 1 (FEN1)

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Area of Science:

  • Molecular Biology
  • DNA Repair Mechanisms
  • Genome Stability

Background:

  • Flap endonuclease 1 (FEN1) is vital for DNA lagging strand maturation and base excision repair (BER).
  • FEN1's role in processing RNA within R-loops and maintaining genome stability is under investigation.
  • Understanding FEN1's RNA processing in R-loops is crucial for elucidating its genome integrity functions.

Purpose of the Study:

  • To investigate how Flap endonuclease 1 (FEN1) processes RNA to resolve R-loops.
  • To examine FEN1's cleavage activity on RNA:DNA hybrid intermediates in R-loops.
  • To determine FEN1's mechanism of action in resolving R-loops via the Base Excision Repair (BER) pathway.

Main Methods:

  • Enzyme activity assays using RNA:DNA hybrid intermediates.
  • Recruitment studies of FEN1 to R-loops in human fibroblasts.
  • Investigating FEN1's interaction with APE1 during R-loop resolution.

Main Results:

  • Human and yeast FEN1 efficiently cleave RNA flaps in R-loop intermediates.
  • FEN1 is recruited to R-loops, with increased recruitment under oxidative DNA damage.
  • FEN1 uses its endonucleolytic activity, coordinating with APE1, to remove RNA from R-loops during BER.

Conclusions:

  • FEN1's endonucleolytic cleavage resolves R-loops through the BER pathway.
  • FEN1 plays a critical role in maintaining genome integrity by resolving R-loops.
  • FEN1 utilizes a unique tracking mechanism to cleave RNA in R-loops, coordinating with BER enzymes.