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The splicing factor XAB2 interacts with ERCC1-XPF and XPG for R-loop processing.

Evi Goulielmaki1, Maria Tsekrekou1,2, Nikos Batsiotos1,2

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The splicing factor XAB2 links RNA splicing and DNA damage response by binding RNA and interacting with repair enzymes. Its release upon DNA damage suggests a role in processing R-loops and maintaining genome stability.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • The interplay between RNA splicing, transcription, and DNA damage response is crucial but not fully understood.
  • Mammalian cells possess intricate mechanisms to maintain genome integrity during these processes.

Purpose of the Study:

  • To elucidate the role of splicing factor XAB2 in linking RNA splicing, transcription, and DNA damage response.
  • To investigate the interaction of XAB2 with the spliceosome and its function in R-loop processing.

Main Methods:

  • In vivo biotinylation tagging in mice to identify XAB2 interactions.
  • XAB2 depletion studies to assess its impact on RNA splicing and DNA damage.
  • Treatment with illudin S and analysis of Csbm/m livers.
  • Immunoprecipitation to study protein-nucleic acid interactions.

Main Results:

  • XAB2 interacts with the spliceosome and binds U4/U6 snRNAs and pre-mRNAs.
  • XAB2 depletion causes intron retention, R-loop formation, and DNA damage.
  • Transcription-blocking DNA lesions release XAB2 from RNA targets.
  • XAB2 forms a complex with ERCC1-XPF and XPG endonucleases, binding RNA:DNA hybrids.

Conclusions:

  • XAB2 functionally connects the spliceosome's response to DNA damage with R-loop processing.
  • This linkage has significant implications for understanding transcription-coupled DNA repair disorders.
  • XAB2 is a key player in maintaining genome stability by integrating splicing and DNA repair pathways.