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Related Experiment Videos

Template definition by Tetrahymena telomerase reverse transcriptase.

M C Miller1, J K Liu, K Collins

  • 1Division of Biochemistry and Molecular Biology, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720-3204, USA.

The EMBO Journal
|August 16, 2000
PubMed
Summary
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Telomerase reverse transcriptase (TERT) has unique N-terminal motifs essential for defining its RNA template. Specific residues within TERT

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Telomerase is a ribonucleoprotein enzyme responsible for extending chromosome ends.
  • It utilizes an integral RNA component as a template for DNA synthesis.
  • The telomerase reverse transcriptase (TERT) protein contains conserved reverse transcriptase (RT) motifs and unique N- and C-terminal extensions.

Purpose of the Study:

  • To investigate the role of Tetrahymena TERT in telomerase activity.
  • To identify TERT regions and residues critical for RNA template definition and utilization.
  • To understand how TERT confers specificity to the telomerase active site.

Main Methods:

  • Site-directed mutagenesis was employed to alter specific residues and motifs within Tetrahymena TERT.

Related Experiment Videos

  • Mutated TERT proteins were analyzed for their impact on telomerase RNP activity.
  • The study focused on template boundary definition and internal template use.

    • Main Results:

    • A novel ciliate-specific motif was identified in the TERT N-terminus, crucial for template definition.
    • Several residues within conserved RT motifs (1, 2, A, and D) were found to be critical for specific aspects of internal template use.
    • TERT residues outside the canonical RT motifs also contribute to telomerase specificity.

    Conclusions:

    • TERT's N-terminal extensions, including a ciliate-specific motif, play a key role in defining the telomerase RNA template.
    • Specific amino acid residues within and beyond the RT motifs of TERT are essential for precise template utilization.
    • The unique specificity of telomerase activity arises from TERT's intricate structure, integrating conserved RT functions with unique extensions.