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

Jennifer A. Fischer, Salvatore J. Caradonna

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

Uracil-DNA glycosylases (UDG/UNG) are enzymes that remove uracil from DNA and initiate baseexcision 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 publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages177-188
Number of pages12
DOIs
StatePublished - Jan 1 2016

Publication series

NameMethods in Molecular Biology
Volume1524
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'Analysis of nuclear uracil DNA-Glycosylase (nUDG) turnover during the cell cycle'. Together they form a unique fingerprint.

  • Cite this

    Fischer, J. A., & Caradonna, S. J. (2016). Analysis of nuclear uracil DNA-Glycosylase (nUDG) turnover during the cell cycle. In Methods in Molecular Biology (pp. 177-188). (Methods in Molecular Biology; Vol. 1524). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6603-5_11