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Functional multimerization of the human telomerase reverse transcriptase.

T L Beattie1, W Zhou, M O Robinson

  • 1Ontario Cancer Institute/Amgen Institute, Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada. tbeattie@ucalgary.ca

Molecular and Cellular Biology
|August 18, 2001
PubMed
Summary
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Human telomerase reverse transcriptase (TERT) forms functional multimers, suggesting a cooperative catalytic core. This multimerization enhances substrate recognition and elongation for telomere maintenance.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Telomerase is a large ribonucleoprotein complex essential for maintaining telomere length.
  • In yeast, telomerase functions as a multimer, but its multimeric state in humans is unknown.
  • Human telomerase reverse transcriptase (hTERT) is a key catalytic subunit.

Purpose of the Study:

  • To investigate whether human TERT (hTERT) forms functional multimers in vivo.
  • To determine the requirements for hTERT multimerization and telomerase activity reconstitution.
  • To explore the functional implications of telomerase multimerization.

Main Methods:

  • Rabbit reticulocyte lysate reconstitution assay.
  • Analysis of human cell extracts.

Related Experiment Videos

  • Complementation assays using catalytically inactive hTERT variants.

    • Main Results:

    • Human TERT (hTERT) forms functional multimers in vitro and in human cell extracts.
    • Catalytically inactive hTERT proteins can complement each other in trans to restore activity.
    • Specific domains of hTERT (N-terminus and RT/C-terminal domains) are required for complementation.
    • Telomerase RNA associates with only one hTERT subunit for activity reconstitution.
    • Telomerase multimerization enhances substrate recognition and elongation.

    Conclusions:

    • The catalytic core of human telomerase likely exists as a functionally cooperative dimer or multimer.
    • Telomerase multimerization is crucial for efficient telomere maintenance.
    • These findings provide insights into the structural and functional organization of human telomerase.