The sporulation-specific enzymes encoded by the DIT1 and DIT2 genes catalyze a two-step reaction leading to a soluble LL-dityrosine-containing precursor of the yeast spore wall.

Dityrosine is a sporulation-specific component of the yeast ascospore wall that is essential for the resistance of the spores to adverse environmental conditions. Dityrosine in vivo exists in both the LL and DL configurations and is part of an insoluble macromolecule of unknown structure. Here we present data indicating that dityrosine of the yeast spore wall is biosynthesized by a different mechanism than dityrosine in other biological systems--e.g., the hard fertilization membrane of the sea urchin egg. We identified two soluble, low molecular weight LL-dityrosine-containing spore wall precursors in extracts of sporulating cells and one precursor containing L-tyrosine. By expression of the previously described sporulation-specific genes DIT1 and DIT2 in vegetative cells, it was shown that DIT1 catalyzes the reaction leading from L-tyrosine to the tyrosine-containing precursor. DIT2, which is a member of the cytochrome P450 superfamily, is responsible for the dimerization reaction leading to the dityrosine-containing precursors. Epimerization of LL- to DL-dityrosine is one of the latest steps in spore wall formation and takes place after the dityrosine-containing precursors are incorporated into the spore wall. On the basis of these findings we suggest a biosynthetic pathway for the top layer of the yeast spore wall.