We induced by UV mutagenesis a series of yeast mutants that were able to utilize dihydroorotic (dhout) and orotic acid (orout) as precursors for pyrimidine biosynthesis. These recessive mutations defined three complementation groups named dhout, orout1 and orout2. The wild-type allele of the gene responsible for dihydroorotate utilization was cloned using the sensitivity of the dhout mutant to 5-fluoroorotate. The DHO gene was sequenced and found to be identical to the URE2 gene. The dhout mutation resulted from the introduction of a stop codon instead of a glutamine at position 59, which led to the production of a truncated Ure2p. Therefore, the URE2 and DHO genes are alleles in yeast.