Selection and characterization of ura5 mutants of Histoplasma capsulatum

The combined use of non-aggregating Histoplasma capsulatum strains and a defined medium which allows quantitative plating of the yeast phase has allowed us to select 5-fluoroorotic acid (5-FOA)-resistant mutants of this dimorphic fungus. Approximately two-thirds of the 5-FOA-resistant strains were auxotrophic for uracil; all were deficient in orotidine-5'-monophosphate pyrophosphorylase (OMPpase) activity. One class of OMPpase mutant (alpha), which retained a low level of OMPpase activity, was auxotrophic in the yeast phase (37 degrees C) but grew slowly in the mycelial phase (25 degrees C) without exogenous uracil. This phenotype was not due to a temperature-sensitive OMPpase activity. Both wild-type and alpha mutants had a higher OMPpase activity in the mycelial phase than the yeast phase; this increased activity may be sufficient to allow mycelial growth of alpha mutants.