Transcript splicing is essential for functional Histoplasma capsulatum URA5 expression

The isolation of auxotrophic markers is important for molecular genetic studies of the dimorphic fungus Histoplasma capsulatum. We have isolated a UV-induced mutant of H. capsulatum, resulting in nonreverting uracil auxotrophy due to a mutation in the URA5 gene. In this study, we show that this mutation is a GG to TA conversion bordering the 5' donor splice site of intron 2. The mutation results in the lack of splicing of intron 2 from the URA5 transcript, and subsequently premature termination of the peptide. This study is the first showing that consensus Group II intron sequences are both utilized and essential for functional expression of a gene in H. capsulatum.