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Programmed translational frameshifting in a gene required for yeast telomere replication

D K Morris1, V Lundblad

  • 1Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA.

Current Biology : CB
|February 21, 1998
PubMed
Summary

Programmed translational frameshifting in yeast produces full-length Est3 protein, essential for telomere replication by telomerase. This mechanism, previously seen in retrotransposons, is crucial for cellular gene function.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Telomeres are critical DNA structures maintained by telomerase, a reverse transcriptase.
  • A genetic screen identified four 'ever shorter telomeres' (EST) genes essential for telomerase function in Saccharomyces cerevisiae.
  • This study focuses on the cloning and characterization of the EST3 gene.

Purpose of the Study:

  • To investigate the function and expression mechanism of the EST3 gene in yeast telomere maintenance.
  • To determine the role of ribosomal frameshifting in EST3 protein production and telomere replication.

Main Methods:

  • Genetic screening to identify telomerase components.
  • Molecular cloning and characterization of the EST3 gene.
  • Analysis of EST3 coding sequence for frameshifting sites.

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  • Mutational analysis of the frameshift site.
  • Immunoblot analysis to detect protein products.

    • Main Results:

    • A +1 ribosomal frameshifting site was identified within the EST3 coding sequence.
    • Mutation of the frameshift site resulted in a loss of telomere replication, similar to an est3 null mutant.
    • Two proteins are synthesized from EST3: a truncated form and the full-length 181 amino-acid Est3 protein.
    • Only the full-length Est3 protein, produced via frameshifting, is required for normal EST3 function.

    Conclusions:

    • Yeast utilizes a programmed translational frameshifting mechanism, akin to retrotransposons, to generate functional Est3 protein.
    • This represents the first documented instance of a cellular gene in yeast employing frameshifting for protein production.
    • A potential link exists between retrotransposition mechanisms and the telomerase pathway for telomere maintenance.