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SFV topoisomerase: sequence specificity in a genetically mapped interval

N Palaniyar1, C Fisher, R Parks

  • 1Department of Molecular Biology and Genetics, University of Guelph, Ontario, Canada.

Virology
|July 15, 1996
PubMed
Summary

Shope fibroma virus (SFV) topoisomerase shares target specificity with vaccinia topoisomerase, recognizing the same DNA motif. However, SFV topoisomerase recognition sites did not significantly impact viral recombination frequencies.

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Area of Science:

  • Virology
  • Molecular Biology
  • Enzymology

Background:

  • Poxviral DNA topoisomerases are sequence-specific enzymes involved in transcription, DNA replication, and recombination.
  • Understanding the relatedness and function of these enzymes is crucial for comprehending poxvirus biology.

Purpose of the Study:

  • To determine the target specificity and catalytic properties of Shope fibroma virus (SFV) topoisomerase.
  • To investigate the influence of SFV topoisomerase on virus-mediated recombination.

Main Methods:

  • SFV topoisomerase was expressed in Escherichia coli and purified as a fusion protein.
  • Catalytic properties and DNA target specificity were assessed.
  • Recombination frequencies were compared across DNA intervals with and without topoisomerase recognition sites.

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Main Results:

  • Recombinant SFV topoisomerase exhibited catalytic properties similar to vaccinia topoisomerase.
  • SFV topoisomerase recognizes the pentanucleotide motif [5'-(C/T)CCTT-3'] and catalyzes DNA relaxation, strand transfer, and cleavage.
  • No significant effect of topoisomerase recognition sites on recombination frequencies was detected, except for a minor stimulation near a poxviral promoter.

Conclusions:

  • Poxviral topoisomerases from different genera share common DNA target specificities.
  • Other enzymatic systems likely mediate the high-frequency recombination observed in poxvirus-infected cells.