The nonpolymorphic, nonclassical class II molecule DM has been shown to be important in the processing and presentation of antigenic epitopes by MHC class II molecules. The dependence of class II molecules on the coexpression of DM varies with the particular allele or epitope studied. In an effort to resolve disparities that exist for some alleles of class II in their requirements for DM, we have constructed a species-matched murine transfection model with which we can test both the functional and biochemical consequences of DM expression for different alleles of murine class II. When we evaluated the ability of class II molecules to form SDS-stable dimer and release CLIP, we find that while I-A(d) requires DM for the formation of SDS-stable dimers and the release of CLIP, I-A(k) is capable of both SDS-stable dimer formation and CLIP release in the absence of DM. Despite the apparent differences in the biochemical consequences of DM expression for I-A(d) and I-A(k), we find that Ag presentation by both alleles can be enhanced by the expression of DM. Interestingly, we find that DM can facilitate the removal of a natural intermediate in Ii processing (p12). The ability of DM to catalyze the dissociation of p12 and possibly larger Ii fragments from class II in vivo offers a possible mechanism to account for the observed DM enhancement of Ag presentation for alleles that have a low affinity for CLIP. Our findings indicate that DM may function in multiple compartments and on multiple class II/Ii substrates.