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

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Abstract
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 DNA (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 corresponding to 4% bromouracil substitution were easily detected. Preliminary density transfer experiments confirm that mtDNA does not replicate in synchrony with nuclear DNA.