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Methods Mol Biol. 2011;761:137-49. doi: 10.1007/978-1-61779-182-6_9.

Analysis of nuclear uracil-DNA glycosylase (nUDG) turnover during the cell cycle.

Author information

1
Department of Molecular Biology, University of Medicine and Dentistry, 08084, Stratford, NJ, USA. fischeje@umdnj.edu

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.

PMID:
21755446
DOI:
10.1007/978-1-61779-182-6_9
[Indexed for MEDLINE]

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