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

Jennifer A. Fischer, Salvatore Caradonna

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationCell Cycle Synchronization
Subtitle of host publicationMethods and Protocols
EditorsGaspar Banfalvi
Pages137-149
Number of pages13
DOIs
StatePublished - Aug 4 2011

Publication series

NameMethods in Molecular Biology
Volume761
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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