The simian varicella virus uracil DNA glycosylase and dUTPase genes are expressed in vivo, but are non-essential for replication in cell culture.
Neurotropic herpesviruses express viral deoxyuridine triphosphate nucleotidohydrolase (dUTPase) and uracil DNA glycosylase (UDG) enzymes which may reduce uracil misincorporation into viral DNA, particularly in neurons of infected ganglia. The simian varicella virus (SVV) dUTPase (ORF 8) and UDG (ORF 59) share 37.7% and 53.9% amino acid identity, respectively, with varicella-zoster virus (VZV) homologs. Infectious SVV mutants defective in either dUTPase (SVV-dUTPase(-)) or UDG (SVV-UDG(-)) activity or both (SVV-dUTPase(-)/UDG(-)) were constructed using recA assisted restriction endonuclease cleavage (RARE) and a cosmid recombination system. Loss of viral dUTPase and UDG enzymatic activity was confirmed in CV-1 cells infected with the SVV mutants. The SVV-dUTPase(-), SVV-UDG(-), and SVV-dUTPase(-)/UDG(-) mutants replicated as efficiently as wild-type SVV in cell culture. SVV dUTPase and UDG expression was detected in tissues derived from acutely infected animals, but not in tissues derived from latently infected animals. Further studies will evaluate the pathogenesis of SVV dUTPase and UDG mutants and their potential as varicella vaccines.
Ward
Toby M
TM
Department of Microbiology and Immunology, 4301 West Markham Street, University of Arkansas for Medical Sciences, Slot 511, Little Rock, AR 72205, United States.
Williams
Marshall V
MV
Traina-Dorge
Vicki
V
Gray
Wayne L
WL
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002644:Chickenpox / Q000821:virology; D002522:Chlorocebus aethiops; D004195:Disease Models, Animal; D015971:Gene Expression Regulation, Enzymologic; D006566:Herpesviridae Infections / Q000821:virology*; D014645:Herpesvirus 3, Human / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D011755:Pyrophosphatases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D051981:Uracil-DNA Glycosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018140:Varicellovirus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
D014764:Viral Proteins; D051981:Uracil-DNA Glycosidase; D011755:Pyrophosphatases; C020020:dUTP pyrophosphatase