title: Analysis of nuclear uracil-DNA glycosylase (nUDG) turnover during the cell cycle.
abstract: Uracil-DNA glycosylases (UDG/UNG) are enzymes that remove uracil from DNA and initiate base-excision repair. These enzymes play a key role in maintaining genomic integrity by reducing the mutagenic events caused by G:C to A:T transition mutations. The recent finding that a family of RNA editing enzymes (AID/APOBECs) can deaminate cytosine in DNA has raised the interest in these base-excision repair enzymes. The methodology presented here focuses on determining the regulation of the nuclear isoform of uracil-DNA glycosylase (nUDG), a 36,000 Da protein. In synchronized HeLa cells, nUDG protein levels decrease to barely detectable levels during the S phase of the cell cycle. Immunoblot analysis of immunoprecipitated or affinity-isolated nUDG reveals ubiquitin-conjugated nUDG when proteolysis is inhibited by agents that block proteasomal-dependent protein degradation.
authors.affiliation: Department of Molecular Biology, University of Medicine and Dentistry, 08084, Stratford, NJ, USA. fischeje@umdnj.edu
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Fischer
authors.forename: Salvatore
authors.initials: S
authors.lastname: Caradonna
mesh_terms: D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology*; D002467:Cell Nucleus / Q000201:enzymology*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D050258:Mitosis Modulators / Q000494:pharmacology; D054875:Ubiquitination / Q000502:physiology; D051981:Uracil-DNA Glycosidase / Q000378:metabolism*
chemical_list: D050258:Mitosis Modulators; D051981:Uracil-DNA Glycosidase