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The canonical RPA complex interacts with Est3 to regulate yeast telomerase activity.

Corinne A Moeller-McCoy1,2, Thomas A Wieser3, Johnathan W Lubin1,2

  • 1Salk Institute for Biological Studies, La Jolla, CA 92037.

Proceedings of the National Academy of Sciences of the United States of America
|February 6, 2025
PubMed
Summary
This summary is machine-generated.

The Est3 subunit of telomerase interacts with the RPA complex, which is crucial for maintaining telomere length in yeast. This interaction ensures telomerase functions correctly, preventing telomere shortening and cellular senescence.

Keywords:
Est3RPARpa2processivitytelomerase

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Telomerase is essential for eukaryotic cell division, maintaining chromosome ends by adding telomeric repeats.
  • In budding yeast, telomerase comprises a catalytic core and regulatory subunits Est1 and Est3.
  • Est1 recruits telomerase to chromosome ends via a telomere-specific complex, but Est3's role was unclear.

Purpose of the Study:

  • To elucidate the regulatory function of the Est3 subunit in telomerase activity.
  • To investigate the interaction between Est3 and the RPA complex in vivo.
  • To understand the mechanism by which RPA influences telomerase function at chromosome ends.

Main Methods:

  • Genetic analysis using mutations in RPA2 and EST3 to observe telomere length phenotypes.
  • Biochemical assays to confirm interactions between Est3 and RPA.
  • Structural analysis of protein interfaces involved in telomerase-RPA binding.

Main Results:

  • Mutations in RPA2 caused an 'Ever shorter telomeres' (Est) phenotype, indicating impaired telomerase function.
  • Est3 interacts with the canonical RPA complex, which is essential for in vivo telomerase function.
  • Compensatory mutations in EST3 and RPA2 support a direct binding interaction.
  • RPA binding to telomerase involves a surface on Est3 analogous to human TPP1.
  • RPA DNA-binding domains are required after RPA-telomerase interaction to stabilize substrate binding and facilitate processivity.

Conclusions:

  • The Est3 subunit's interaction with RPA is critical for telomerase function and telomere maintenance.
  • RPA plays a dual role: recruiting telomerase and stabilizing its interaction with telomeric DNA for processive action.
  • Yeast telomerase utilizes two distinct ssDNA-binding complexes, highlighting the importance of single-stranded DNA management in telomere biology.