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Related Experiment Videos

Telomerase is processive.

C W Greider1

  • 1Cold Spring Harbor Laboratory, New York 11724.

Molecular and Cellular Biology
|September 1, 1991
PubMed
Summary
This summary is machine-generated.

Telomerase synthesizes long DNA repeats processively using an RNA template. The enzyme

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Telomerase is a ribonucleoprotein enzyme responsible for maintaining telomere length.
  • It synthesizes tandem DNA repeats using an intrinsic RNA template.
  • The specific repeat sequence synthesized is d(TTGGGG).

Purpose of the Study:

  • To investigate the processivity and elongation mechanism of telomerase.
  • To characterize the synthesis of d(TTGGGG) repeats by telomerase.
  • To understand the role of the RNA template in telomerase activity.

Main Methods:

  • In vitro telomerase assays were performed using d(TTGGGG)n primer oligonucleotides.
  • Product length and synthesis rates were analyzed.
  • Dilution and primer challenge experiments assessed processivity.

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  • Banding pattern analysis revealed reaction pauses and product characteristics.

    • Main Results:

    • Telomerase synthesized long products (>2,000 nucleotides) processively within 60 minutes.
    • Quantification showed approximately 520 nucleotides synthesized before half-enzyme dissociation.
    • A characteristic 6-nucleotide banding pattern was observed, with a pause after the first G.
    • Processivity and banding patterns suggest a translocation step in the elongation mechanism.

    Conclusions:

    • Telomerase exhibits significant processivity in synthesizing d(TTGGGG) repeats.
    • The enzyme reinitiates synthesis on new primers after dissociation.
    • Elongation mechanism likely involves a translocation step following template copying.
    • Understanding telomerase mechanism is crucial for telomere biology and cancer research.