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PMID: 8223452 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

dUTP pyrophosphatase is an essential enzyme in Saccharomyces cerevisiae.

The EMBO journal ·Vol. 12 ·No. 11 ·1993-11-00 ·Pages 4425-31

Gadsden MH, McIntosh EM, Game JC, Wilson PJ, Haynes RH

Abstract

dUTP pyrophosphatase (dUTPase; EC 3.6.1.23) catalyses the hydrolysis of dUTP to dUMP and PPi and thereby prevents the incorporation of uracil into DNA during replication. Although it is widely believed that dUTPase is essential for cell viability because of this role, direct evidence supporting this assumption has not been presented for any eukaryotic system. We have analysed the role of dUTPase (DUT1) in the life cycle of yeast. Using gene disruption and tetrad analysis, we find that DUT1 is necessary for the viability of S. cerevisiae; however, under certain conditions dut1 null mutants survive if supplied with exogenous thymidylate (dTMP). Analyses with isogenic uracil-DNA-glycosylase (UNG1) deficient or proficient strains indicate that in the absence of dUTPase, cell death results from the incorporation of uracil into DNA and the attempted repair of this damage by UNG1-mediated excision repair. However, in dut1 ung1 double mutants, starvation for dTMP causes dividing cells to arrest and die in all phases of the cell cycle. This latter effect suggests that the extensive stable substitution of uracil for thymine in DNA leads to a general failure in macromolecular synthesis. These results are in general agreement with previous models in thymine-less death that implicate dUTP metabolism. They also suggest an alternative approach for chemotherapeutic drug design.

Related Genes
DUT1
MeSH Terms
Amino Acid Sequence Base Sequence Cell Cycle/physiology Chromosome Mapping Chromosomes, Fungal Cloning, Molecular Crosses, Genetic DNA Glycosylases DNA Repair Genes, Fungal Genes, Lethal Molecular Sequence Data Mutagenesis, Insertional Mutation N-Glycosyl Hydrolases/genetics Pyrophosphatases/genetics Saccharomyces cerevisiae/enzymology,genetics Sequence Analysis, DNA Sequence Homology, Amino Acid Thymidine Monophosphate/metabolism Thymine/metabolism Uracil-DNA Glycosidase
Chemicals
Thymidine Monophosphate DNA Glycosylases N-Glycosyl Hydrolases Uracil-DNA Glycosidase Pyrophosphatases dUTP pyrophosphatase Thymine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Gadsden M H
Department of Biology, York University, Toronto, Ontario, Canada.
McIntosh E M
Game J C
Wilson P J
Haynes R H
References (42)
42 references, click to expand
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1993-11-00
Pages
4425-31
Language
English
Region
England
NLM ID
8208664
PMCID
PMC413740
Subset
IM
Grants
NIGMS NIH HHS · GM30990 · United States
Databases
GENBANK
X74263
External Links
PubMed source PMC Full Text View journal details
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