Identification of a poxvirus gene encoding a uracil DNA glycosylase

Chris Upton, David T. Stuart, Douglas McFadden

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

An open reading frame, BamHI D6R, from the central highly conserved region of the Shope fibroma virus (SFV) genome was sequenced and found to have significant homology to that of uracil DNA glycosylases from a number of organisms. Uracil DNA glycosylase catalyzes the initial step in the repair pathway that removes potentially mutagenic uracil from duplex DNA. The D6R polypeptide was expressed in reticulocyte lysates programed with RNA transcribed from an expression vector containing the T7 RNA polymerase pro-moter. A highly specific ethidium bromide fluorescence assay of the in vitro translation product determined that the encoded protein does indeed possess uracil DNA glycosylase activity. Open reading frames from other poxviruses, including vaccinia virus (HindIII D4R) and fowlpox (D4), are highly homologous to D6R of SFV and are predicted to encode uracil DNA glycosylases. Identification of the SFV uracil DNA glycosylase provides evidence that this poxviral protein is involved in the repair of the viral DNA genome. Since this enzyme performs only the initial step required for the removal of uracil from DNA, creating an apyrimidinic site, we suggest that other, possibly virus-encoded, repair activities must be present in the cytoplasm of infected cells to complete the uracil excision repair pathway.

Original languageEnglish (US)
Pages (from-to)4518-4522
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number10
DOIs
StatePublished - May 15 1993
Externally publishedYes

Fingerprint

Uracil-DNA Glycosidase
Poxviridae
Rabbit Fibroma Virus
Uracil
Genes
Open Reading Frames
Fowlpox
Ethidium
Vaccinia virus
Viral Genome
Reticulocytes
DNA
Viral DNA
DNA Repair
Cytoplasm
Proteins
Fluorescence
Genome
RNA
Viruses

Keywords

  • DNA repair
  • Ethidium bromide fluorescence assay
  • Shope fibroma virus
  • Vaccinia virus

ASJC Scopus subject areas

  • General

Cite this

Identification of a poxvirus gene encoding a uracil DNA glycosylase. / Upton, Chris; Stuart, David T.; McFadden, Douglas.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, No. 10, 15.05.1993, p. 4518-4522.

Research output: Contribution to journalArticle

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