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Related Experiment Videos

Accuracy of DNA primase.

S S Zhang1, F Grosse

  • 1Max-Planck-Institute for Experimental Medicine, Department of Chemistry, Göttingen, F.R.G.

Journal of Molecular Biology
|December 5, 1990
PubMed
Summary

DNA primase exhibits low base-pairing fidelity, making it the least accurate DNA-synthesizing enzyme known. This inaccuracy highlights the necessity of a priming mechanism for DNA synthesis initiation.

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Area of Science:

  • Molecular Biology
  • Enzymology
  • Biochemistry

Background:

  • DNA primase synthesizes RNA primers essential for DNA replication initiation.
  • Understanding the fidelity of DNA primase is crucial for comprehending DNA synthesis accuracy.

Purpose of the Study:

  • To investigate the base-pairing fidelity of calf thymus DNA primase in vitro.
  • To quantify the misinsertion frequency of nucleotides by DNA primase.

Main Methods:

  • Utilized a misinsertion assay with synthetic polydeoxynucleotide templates.
  • Employed in vitro studies to assess nucleotide incorporation by DNA primase.

Main Results:

  • DNA primase demonstrated low fidelity, with GMP misinsertions occurring at a frequency of 1/1600 on a poly(dT) template.
  • UMP and CMP misinsertion frequencies were 1/300 and 1/500, respectively, on a poly(dT) template.
  • On a poly(dC, dT) template, UMP and CMP misinsertion frequencies were 1/200 and 1/300, respectively.

Conclusions:

  • Calf thymus DNA primase is the least accurate polynucleotide-synthesizing enzyme identified to date.
  • The observed low fidelity underscores the critical role of a priming mechanism in initiating DNA synthesis.
  • Further research may explore mechanisms to enhance DNA primase accuracy or compensate for its errors.

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