Linking uracil base excision repair and 5-fluorouracil toxicity in yeast

Lauren Seiple, Pawel Jaruga, Miral Dizdaroglu, James Stivers

Research output: Contribution to journalArticle

Abstract

5-fluorouracil (5-FU) is a widely used anticancer drug that disrupts pyrimidine nucleotide pool balances and leads to uracil incorporation in DNA, which is then recognized and removed by the uracil base excision repair (BER) pathway. Using complementary biochemical and genetic approaches we have examined the role of uracil BER in the cell killing mechanism of 5-FU. A yeast strain lacking the enzyme uracil DNA glycosylase (Ung1), which excises uracil from the DNA backbone leaving an abasic site, showed significant protection against the toxic effects of 5-FU, a G1/S cell cycle arrest phenotype, and accumulated massive amounts of U/A base pairs in its genome (∼4% of T/A pairs were now U/A). A strain lacking the major abasic site endonuclease of Saccharomyces cerevisiae (Apn1) showed significantly increased sensitivity to 5-FU with G2/M arrest. Thus, efficient processing of abasic sites by this enzyme is protective against the toxic effects of 5-FU. However, contrary to expectations, the Apn1 deficient strain did not accumulate intact abasic sites, indicating that another repair pathway attempts to process these sites in the absence Apn1, but that this process has catastrophic effects on genome integrity. These findings suggest that new strategies for chemical intervention targeting BER could enhance the effectiveness of this widely used anticancer drug.

Original languageEnglish (US)
Pages (from-to)140-151
Number of pages12
JournalNucleic Acids Research
Volume34
Issue number1
DOIs
StatePublished - Jan 2006

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Uracil
Fluorouracil
DNA Repair
Yeasts
Poisons
Uracil-DNA Glycosidase
Pyrimidine Nucleotides
Genome
G1 Phase Cell Cycle Checkpoints
Endonucleases
DNA
Enzymes
Base Pairing
Pharmaceutical Preparations
Saccharomyces cerevisiae
Molecular Biology
Phenotype

ASJC Scopus subject areas

  • Genetics

Cite this

Linking uracil base excision repair and 5-fluorouracil toxicity in yeast. / Seiple, Lauren; Jaruga, Pawel; Dizdaroglu, Miral; Stivers, James.

In: Nucleic Acids Research, Vol. 34, No. 1, 01.2006, p. 140-151.

Research output: Contribution to journalArticle

Seiple, Lauren ; Jaruga, Pawel ; Dizdaroglu, Miral ; Stivers, James. / Linking uracil base excision repair and 5-fluorouracil toxicity in yeast. In: Nucleic Acids Research. 2006 ; Vol. 34, No. 1. pp. 140-151.
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