Specific DNA-labelling by exogenous thymidine-5′-monophosphate in Saccharomyces cerevisiae

Fäth, Wolfgang W.; Brendel, Martin

doi:10.1007/bf00269387

Cited by 21 publications

(5 citation statements)

References 15 publications

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“…At about this time the problem of specific radiolabeling of DNA in S. cerelisiaie was solved by the isolation of strains that can take up thymidine monophosphate and incorporate it into their genome (25,48,105,287) and later by the isolation of authentic thymidine monophosphate auxotrophs (27,48,145). However, the use of these mutants for measuring excision of pyrimidine dimers would require that the thymidine monophosphate-defective genotype be systematically introduced into all RAD3 epistasis group mutants, a labor-intensive task that may not be compatible with viability in all cases (for example, there are indications that a rad6 strain that can take up deoxythymidine monophosphate is inviable [M. Brendel, unpublished observations]).…”

Section: Characterization Of Defective Dna Repairmentioning

confidence: 99%

Deoxyribonucleic acid repair in the yeast Saccharomyces cerevisiae.

Friedberg¹

1988

Microbiological Reviews

179

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Section: Characterization Of Defective Dna Repairmentioning

confidence: 99%

Deoxyribonucleic acid repair in the yeast Saccharomyces cerevisiae.

Friedberg¹

1988

Microbiological Reviews

179

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“…cerevisiae strain T6-425 was grown with shaking at 37°C in medium N of Fath and Brendel (1974;1976) because it grows satisfactorily only in this medium. Exponential growth was established before the addition of cytosine or 5FC to a final concentration of 100 mg/litre.…”

mentioning

confidence: 99%

“…Polak and Scholer (1975) have shown that these mechanisms apply to C. albicans. One mutant strain of S. cerevisiae, T6-425, has been found to utilise thymidine (Fath and Brendel, 1974;1976) and we used it to investigate the effect of 5FC on the incorporation of thymidine into DNA as a measure of DNA synthesis. Tritiated thymidine monophosphate (lpCi/ml) was added to cultures of S. cerevisiae strain T6-425 containing cytosine or 5FC.…”

mentioning

confidence: 99%

The Effect of 5-Fluorocytosine on the Blastospores and Hyphae of Candida Albicans

Polak¹,

Wain

1979

Journal of Medical Microbiology

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5-FLUOROCYTOSINE (5FC) has an important place in the treatment of systemic mycoses in man. Its mode of action was initially explained by an extensive incorporation of the deaminated form, 5-fluorouracil, into fungal RNA, with this leading to a disturbance of the internal amino-acid pool (Polak, 1974). Recently, the inhibition of RNA and DNA syntheses has been reported by Polak and Wain, (1977; and by Diasio, Bennett and Myers (1978). The inhibition of DNA synthesis in Escherichia coli by trimethoprim or by 5-fluorodeoxyuridine is rapidly bactericidal (Cohen et al., 1958 ;Angehrn and Then, 1973) but there is no evidence of 5FC having a rapid fungicidal action on Candida albicans (Scholer and Polak, 1973).Primary resistance to 5FC has been reported by Speller and Davies (1973) and is considered to involve several mechanisms in the pathway of RNA synthesis (Polak and Scholer, 1975), which mechanisms may also affect DNA synthesis. The dimorphism of C. albicans affects the development of resistance to 5FC (Davies and Savage, 1974) and the hyphal form may be associated with the invasive ability of this opportunist fungus (Montes and Wilborn, 1968 ;Cawson and Rajasingham, 1972; Wain, Price and Cawson, 1976~).Because the inhibition of DNA synthesis by 5FC was accompanied by an increase in blastospore cell volume and hyphal length, the effect of the drug on the protein and carbohydrate metabolism of the fungus in its hyphal and blastospore forms has been investigated and studies with radioactive precursors have been undertaken to elucidate the inhibition of DNA and RNA syntheses. MATERIALS AND METHODS Microorganisms

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“…Consequently, yeast DNA has been labeled with other radioactive bases or nucleotides with the attendant problem of incorporation of label into ribonucleic acid. To overcome this problem, mutants have been isolated (1,2,5,6,15,20) after selection for growth when the de novo pathway for deoxythymidine 5'-monophosphate synthesis was blocked through the combined action of aminopterin, an inhibitor ofdihydrofolate reductase, and sulfanilamide, an inhibitor of folate synthesis (1,15,20). The use of these mutants has some drawbacks.…”

mentioning

confidence: 99%

“…The use of these mutants has some drawbacks. First, high levels of exogenous thymidine 5'-monophosphate (TMP) are necessary for optimal incorporation, presumably, because a phosphatase degrades TMP to thymidine andinorganic phosphate (5). Secondly, perhaps as a consequence of the selection procedure (2), the mutants produce a high percentage of petite colonies, which render them impractical for ex-periments dealing with wild-type mitochondrial DNA (mtDNA).…”

mentioning

confidence: 99%

Bromodeoxyuridine 5'-monophosphate incorporation into yeast nuclear and mitochondrial deoxyribonucleic acid

Leff¹,

Lam²

1976

J Bacteriol

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Standard laboratory yeast strains can be enriched for thymidine 5'-monophosphate (TMP) uptake derivatives that generate only a low percentage of respiratory-deficient colonies (petites) under inhibition of TMP biosynthesis. Such mutants incorporated bromodeoxyuridine 5'-monophosphate (BrdUMP) into both nuclear and mitochondrial deoxyribonucleic acid (mtDNA); however, they showed a selectivity for TMP over BrdUMP incorporation. The preferential incorporation of [3H]TMP or BrdUMP into mtDNA was strain dependent. The density increments after growth in the presence of BrdUMP reached 50 mg/ml for nuclear DNA and 22 mg/ml for mtDNA in CsCl gradients. Density shifts correspondisng to 4% bromouracil substitution were easily detected. Preliminary density transfer experiments confirm that mtDNA does not replicate in synchrony with nuclear DNA.

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Specific DNA-labelling by exogenous thymidine-5′-monophosphate in Saccharomyces cerevisiae

Cited by 21 publications

References 15 publications

Deoxyribonucleic acid repair in the yeast Saccharomyces cerevisiae.

Deoxyribonucleic acid repair in the yeast Saccharomyces cerevisiae.

The Effect of 5-Fluorocytosine on the Blastospores and Hyphae of Candida Albicans

Bromodeoxyuridine 5'-monophosphate incorporation into yeast nuclear and mitochondrial deoxyribonucleic acid

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