Molecular mechanism of base excision repair of uracil-containing DNA in yeast cell-free extracts

Zhigang Wang, Xiaohua Wu, Errol C. Friedberg

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Base excision repair (BER) constitutes a ubiquitous excision repair mechanism, which is responsible for the removal of multiple types of damaged and inappropriate bases in DNA. We have employed a yeast cell-free system to examine the biochemical mechanism of the BER pathway in lower eukaryotes. Using uracil-containing DNA as a model substrate, we demonstrate that yeast BER requires Apn1 protein, an Escherichia coli endonuclease IV homolog. In extracts of an apn1 deletion mutant, the 5'-incision at AP (apurinic/apyrimidinic) sites is not detectable, supporting the notion that yeast contains only one major 5'-AP endonuclease. The processing of the 5'- deoxyribose phosphate moieties was found to be a rate-limiting step. During BER of uracil-containing DNA, repair patch sizes of 1-5 nucleotides were detected, with single nucleotide repair patches predominant.

Original languageEnglish (US)
Pages (from-to)24064-24071
Number of pages8
JournalJournal of Biological Chemistry
Volume272
Issue number38
DOIs
StatePublished - Sep 19 1997

Fingerprint

Uracil
Cell Extracts
DNA Repair
Yeast
Repair
Yeasts
Cells
DNA
Deoxyribonuclease IV (Phage T4-Induced)
Nucleotides
DNA-(Apurinic or Apyrimidinic Site) Lyase
Deoxyribose
Cell-Free System
Escherichia coli Proteins
Eukaryota
Endonucleases
Phosphates
Substrates
Processing

ASJC Scopus subject areas

  • Biochemistry

Cite this

Molecular mechanism of base excision repair of uracil-containing DNA in yeast cell-free extracts. / Wang, Zhigang; Wu, Xiaohua; Friedberg, Errol C.

In: Journal of Biological Chemistry, Vol. 272, No. 38, 19.09.1997, p. 24064-24071.

Research output: Contribution to journalArticle

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