Orf virus encodes a functional dUTPase gene.
The present study is the first report on the functional activity of a parapoxvirus-encoded dUTPase. The dUTPase gene of the attenuated orf virus (ORFV), strain D1701, was expressed as a bacterial thioredoxin fusion protein. In vitro assays showed that ORFV dUTPase was highly specific for dUTP as substrate. The enzyme was active over a broad pH range (pH 6.0-9.0), with maximal enzymatic activity at pH 7.0 in the presence of Mg(2+) cations. Kinetic studies of the recombinant ORFV dUTPase revealed an apparent K(m) of 4.0 microM, which is more similar to that of the mammalian or African swine fever virus enzyme than to the K(m) of vaccinia virus dUTPase. Enzyme activity was also found with purified ORFV particles, indicating its virion association.
Cottone
R
R
Federal Research Centre for Virus Diseases of Animals, Institute for Immunology, Paul-Ehrlich-Straße 28, D-72076 Tübingen, Federal Republic of Germany1.
Büttner
M
M
Federal Research Centre for Virus Diseases of Animals, Institute for Immunology, Paul-Ehrlich-Straße 28, D-72076 Tübingen, Federal Republic of Germany1.
McInnes
C J
CJ
Moredun Research Institute, International Research Centre, Pentlands Science Park, Penicuik, Midlothian EH26 OPZ, UK2.
Wood
A R
AR
Moredun Research Institute, International Research Centre, Pentlands Science Park, Penicuik, Midlothian EH26 OPZ, UK2.
Rziha
H-J
HJ
Federal Research Centre for Virus Diseases of Animals, Institute for Immunology, Paul-Ehrlich-Straße 28, D-72076 Tübingen, Federal Republic of Germany1.
D003856:Deoxyuracil Nucleotides / Q000378:metabolism; D009923:Orf virus / Q000201:enzymology* / Q000235:genetics; D011755:Pyrophosphatases / Q000235:genetics*
D003856:Deoxyuracil Nucleotides; C027078:deoxyuridine triphosphate; D011755:Pyrophosphatases; C020020:dUTP pyrophosphatase