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Related Experiment Videos

Characterization of human telomerase complex

S Ramakrishnan1, H W Sharma, A D Farris

  • 1Department of Oncology, Hoffmann-La Roche, 340 Kingsland Street, Nutley, NJ 07110, USA.

Proceedings of the National Academy of Sciences of the United States of America
|September 18, 1997
PubMed
Summary
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Researchers identified a bioinformatics method to enrich and characterize human telomerase, a complex enzyme crucial for DNA replication. This study reveals evolutionary conservation of telomerase proteins, linking the autoantigen Ro60 to telomerase activity.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genetics

Background:

  • Telomerase is a ribonucleoprotein complex responsible for adding telomeric repeats to chromosomal DNA ends.
  • Mammalian telomerase characterization is challenging due to low expression levels.

Purpose of the Study:

  • To develop a bioinformatics approach for enriching and characterizing the human telomerase complex.
  • To investigate potential evolutionary conservation of telomerase proteins.

Main Methods:

  • Utilized local sequence homology search to identify similarities between Tetrahymena p80 and human autoantigen Ro60.
  • Employed antibodies against Ro60 for immunoprecipitation of telomerase activity.
  • Analyzed immunoprecipitated material using native gel electrophoresis and in vitro binding assays with a TTAGGG probe.

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Main Results:

  • Detected similarity between Tetrahymena p80 and Ro60, with Ro60 antibodies precipitating telomerase activity.
  • Confirmed cross-recognizability between Ro60 and p80 proteins.
  • Localized telomerase activity and its RNA component to a specific doublet, demonstrating RNA template-dependent binding to a TTAGGG probe.

Conclusions:

  • The findings suggest an evolutionary conservation of telomerase proteins across species.
  • The autoantigen Ro60 is implicated in human telomerase complex and activity.
  • The developed bioinformatics approach facilitates telomerase enrichment and characterization.