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Telomerase Repairs Collapsed Replication Forks at Telomeres.

Samah Matmati1, Sarah Lambert2, Vincent Géli1

  • 1Marseille Cancer Research Centre (CRCM), U1068 INSERM, UMR7258 CNRS, UM105 Aix-Marseille University, Institut Paoli-Calmettes, Ligue Nationale Contre le Cancer (équipe labellisée) Marseille, F-13009, France.

Cell Reports
|March 12, 2020
PubMed
Summary

Replication of telomeres is impaired in fission yeast lacking telomerase, leading to stalled forks. Homologous recombination and telomerase compete for DNA ends, with telomerase shielding telomeres from recombination.

Keywords:
homologous recombinationreplication stresssenescencetelomerasetelomere

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Telomeres are crucial for chromosome stability but challenging to replicate.
  • Replication stress at telomeres can threaten genome integrity.
  • Telomerase is essential for telomere maintenance in many organisms.

Purpose of the Study:

  • To investigate telomere replication dynamics in fission yeast lacking telomerase.
  • To identify mechanisms that maintain telomere integrity during replication stress.
  • To understand the interplay between telomerase, Ku, and homologous recombination at telomeres.

Main Methods:

  • Two-dimensional gel electrophoresis to analyze replication intermediates.
  • Investigating the roles of Rad51, MRN complex, Ctp1, Ku, and telomerase.
  • Utilizing telomerase-negative and catalytically dead telomerase mutant fission yeast strains.

Main Results:

  • Telomere replication is severely impaired, with accumulation of stalled and collapsed replication forks.
  • Homologous recombination factors (Rad51, MRN, Ctp1) are critical for processing these intermediates and facilitating fork restart.
  • A catalytically dead telomerase mutant impairs Ku recruitment to telomeres.
  • Telomerase and Ku compete for binding to telomeric free DNA ends, likely originating from reversed forks.

Conclusions:

  • Homologous recombination is essential for processing stalled and collapsed telomeric replication forks in the absence of telomerase.
  • Telomerase and Ku compete for telomeric DNA ends, suggesting a mechanism for telomere protection.
  • Ku removal at collapsed forks allows telomerase to repair broken telomeres, preventing excessive homologous recombination and maintaining telomere integrity.