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RNAPII-dependent ATM signaling at collisions with replication forks.

Elias Einig1, Chao Jin1, Valentina Andrioletti1,2

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Oncogenic signaling causes transcription-replication conflicts (TRCs), leading to DNA damage. This study shows TRCs activate ATM kinase for DNA repair, with WRNIP1 regulating this process.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Deregulation of RNA Polymerase II (RNAPII) by oncogenic signaling causes transcription-replication conflicts (TRCs).
  • TRCs can lead to DNA damage and genomic instability in cancer cells.
  • ATM kinase activation is crucial for DNA repair pathways.

Purpose of the Study:

  • To investigate the role of RNAPII in nucleating ATM kinase activation at TRCs.
  • To elucidate the function of WRNIP1 and HUWE1 in regulating ATM activation during TRCs.
  • To understand the cellular response to replicative stress.

Main Methods:

  • Investigated RNAPII behavior and ATM kinase activation at TRCs using cellular and molecular biology techniques.
  • Utilized genetic mutations in HUWE1 to study its effect on WRNIP1 binding and ATM activation.
  • Employed hydroxyurea treatment to induce TRCs and assess WRNIP1 translocation and ATM activation.

Main Results:

  • Elongating RNAPII nucleates ATM kinase activation at TRCs to promote DNA repair.
  • The ATPase WRNIP1 associates with RNAPII and limits ATM activation.
  • HUWE1's ubiquitin ligase activity is required for WRNIP1 binding to RNAPII; its mutation disrupts this interaction, leading to increased ATM activation.
  • TRCs and WRNIP1 translocation are rapidly induced by hydroxyurea, activating ATM for DNA repair.

Conclusions:

  • TRCs serve as a platform for ATM kinase activation, facilitating DNA repair.
  • WRNIP1 acts as a negative regulator of ATM activation at TRCs, requiring HUWE1 activity.
  • TRCs and WRNIP1 translocation represent a controlled mechanism for stalling replication forks and activating ATM in response to replicative stress.