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Related Experiment Videos

Orf virus encodes a functional dUTPase gene.

R Cottone1, M Büttner1, C J McInnes2

  • 1Federal Research Centre for Virus Diseases of Animals, Institute for Immunology, Paul-Ehrlich-Straße 28, D-72076 Tübingen, Federal Republic of Germany1.

The Journal of General Virology
|April 19, 2002
PubMed
Summary

This study reveals the functional activity of orf virus (ORFV) dUTPase, an enzyme crucial for DNA replication. The ORFV dUTPase enzyme shows high substrate specificity and is associated with viral particles.

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Area of Science:

  • Virology
  • Enzymology
  • Molecular Biology

Background:

  • Parapoxviruses encode dUTPase enzymes essential for DNA metabolism.
  • Understanding viral dUTPase function is key to comprehending viral replication strategies.

Purpose of the Study:

  • To characterize the functional activity of a parapoxvirus-derived dUTPase.
  • To investigate the biochemical properties and substrate specificity of the orf virus (ORFV) dUTPase.

Main Methods:

  • Expression of the ORFV dUTPase gene as a bacterial thioredoxin fusion protein.
  • In vitro enzymatic assays to determine substrate specificity, optimal pH, and kinetic parameters.
  • Analysis of dUTPase activity in purified ORFV particles.

Main Results:

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  • The recombinant ORFV dUTPase demonstrated high specificity for dUTP.
  • Optimal enzyme activity was observed at pH 7.0 in the presence of Mg(2+) ions.
  • Kinetic analysis revealed an apparent K(m) of 4.0 microM, comparable to mammalian and African swine fever virus dUTPases.
  • Enzyme activity was detected in purified ORFV particles, suggesting virion association.

Conclusions:

  • The study provides the first report on the functional characterization of a parapoxvirus-encoded dUTPase.
  • ORFV dUTPase exhibits biochemical properties distinct from vaccinia virus dUTPase.
  • The virion association of ORFV dUTPase suggests a potential role in viral DNA replication or genome integrity within the virion.