Mutants for the specific labelling of DNA in Saccharomyces cerevisiae

Jannsen, Sigrid; Witte, Irene; Megnet, Roland

doi:10.1016/0005-2787(73)90243-8

Cited by 28 publications

(10 citation statements)

References 4 publications

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“…Thus, strains carrying cdc2l can be starved for dTMP simply by incubating them at 360C. Game (19) has shown that cdc2l strains incubated at the restrictive temperature gradually lose viability, and the kinetics of cell killing are similar to those displayed by other dTMPdeprived thymidylate auxotrophs (12,14 21 involved expression of the ura3 allele (situated on the left arm of chromosome V). Among these uracil requirers, 12 colonies were found that also were auxotrophic for one or more markers on the right arm of chromosome V. In some instances the expressed alleles were originally in a trans configuration (e.g., two colonies were ura-arg-trp-), whereas in other cases they were in a cis configuration (e.g., five colonies were cans ura -horn-ilv-).…”

Section: Fig 3 Fdump-induced Expression Of Cycloheximide Resistancementioning

confidence: 99%

“…The in vitro activity of thymidylate synthetase from S. cerevisiae requires N5,N'0-methylenetetrahydrofolate (17). Concurrent treatment with aminopterin and sulfanilamide inhibits thymidylate biosynthesis (I3, 15,[21][22][23][24][25], presumably by depleting endogenous tetrahydrofolate pools (Fig. 1).…”

Section: Fig 3 Fdump-induced Expression Of Cycloheximide Resistancementioning

confidence: 99%

“…In general, the dihydrofolate (FH2) analogue aminopterin inhibits dihydrofolate reductase (reaction 2) whereas sulfanilamide, an analogue of p-aminobenzoate, poisons de novo folate synthesis (reaction 3) (29). In yeast, treatment with aminopterin plus sulfanilamide blocks dTMP biosynthesis (13,15,(21)(22)(23)(24)(25), presumably by removing FH4 and so reducing the level of N 5,N'0-methylenetetrahydrofolate. dTMP (tup-) were sensitive to FdUMP whereas S288C, a wild-type strain (TUP+), was not.…”

mentioning

confidence: 99%

“…Drugs that inhibit this enzyme or that limit the supply of reduced folates can abolish dTMP biosynthesis. In yeast, the combination of aminopterin plus sulfonamide is a potent inhibitor (13,15,(21)(22)(23)(24)(25). Cells thus treated require thymidylate, adenine, histidine, and methionine for growth (13,15 Previous work had shown that dTMP deprivation in yeast was not mutagenic for nuclear genes, but caused mitochondrial point mutations and cytoplasmic petites (14).…”

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confidence: 99%

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Induction of mitotic recombination in yeast by starvation for thymine nucleotides.

Kunz¹,

Barclay²,

Game³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

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The biosynthesis of thymine nucleotides in Saccharomyces cerevisiae can be inhibited either by genetic lesions in the structural gene for thymidylate synthetase (TMP1) or by drugs that prevent the methylation of dUMP to dTMP. This methylation can be blocked by folate antagonists. We find that 5-fluoro-dUMP (FdUMP) is also an effective inhibitor in vivo. Inhibition of dTMP biosythesis by these three different routes causes thymineless death. In addition to being cytotoxic, we find that FdUMP is highly recombinagenic in yeast but does not induce nuclear gene mutations. Provision of exogenous dTMP eliminates this induced mitotic recombination and cell killing. Similar results were obtained when a thymineless condition was provoked in cells by antifolate drugs or by dTMP deprivation in strains auxotrophic for this nucleotide. These findings show that, in contrast to the situation in prokaryotes, starvation for thymine nucleotides in yeast induces genetic recombination but is not mutagenic.

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Section: Fig 3 Fdump-induced Expression Of Cycloheximide Resistancementioning

confidence: 99%

Section: Fig 3 Fdump-induced Expression Of Cycloheximide Resistancementioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Induction of mitotic recombination in yeast by starvation for thymine nucleotides.

Kunz¹,

Barclay²,

Game³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

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“…It has been possible to isolate recessive dTMP uptake (tup) mutants that permit the incorporation of dTMP into DNA. These mutants fall into at least four complementation groups (2,5,8). One of these mutations, tupl, is pleiotropic, also depressing both the ability of MATa strains to mate and the detection of canavanine-resistant mutants induced by UV light (6,8).…”

mentioning

confidence: 99%

Suppressor of deoxythmidine monophosphate uptake in Saccharomyces cerevisiae

Remer¹,

Sherman²,

Kraig³

et al. 1979

J Bacteriol

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Although yeast cannot normally incorporate exogenous deoxythymidine 5'monophosphate (dTMP) into deoxyribonucleic acid, mutants able to do so have been isolated. We have characterized a recessive suppressor of dTMP uptake (sotl) that prevents strains carrying either tupi, tup2, or tup4 from growing on selective medium. The sotl mutation maps between radl and the centromere of chromosome XVI, and is unlinked to any of the tup mutations. The sotl mutation does not suppress the other pleiotropic effects of the tupl mutant, notably the lack of mating of tupl MATa strains. The sotl mutation specifically blocks the uptake of dTMP into tup strains. After growing a sotl strain in medium containing [3H]dTMP, we showed that the medium still contained more than 90% of the original [3H]dTMP and that this medium could support the incorporation of [3H]dTMP by a tup2 strain. Therefore, sotl strains do not degrade dTMP in the medium. The sotl mutation had no effect on the uptake of other nutrients essential for growth, including several amino acids, adenine, and uracil.

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Genetic effects of deoxyribonucleotide pool imbalances

Kunz

1982

Environ. mutagen

124

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In vivo DNA precursor imbalances can have profound genetic consequences in prokaryotes and eukaryotes. In vitro studies have demonstrated that DNA replication fidelity is dependent on correct balances of deoxyribonucleoside triphosphates during DNA synthesis. These findings suggest that intracellular concentrations of DNA precursors may be regulated in order to minimize the frequency of genetic change. In addition, they indicate the existence of important nonDNA targets for the induction of mutation, recombination, chromosome aberrations, etc. In this article, the genetic effects of deoxyribonucleotide pool imbalances are reviewed.

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Mutants for the specific labelling of DNA in Saccharomyces cerevisiae

Cited by 28 publications

References 4 publications

Induction of mitotic recombination in yeast by starvation for thymine nucleotides.

Induction of mitotic recombination in yeast by starvation for thymine nucleotides.

Suppressor of deoxythmidine monophosphate uptake in Saccharomyces cerevisiae

Genetic effects of deoxyribonucleotide pool imbalances

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