FIGURE 5.

Ubiquitin is attached to multiple residues on mDia2. A, ClustalW protein sequence alignment of mouse mDia1, human mDia1, mouse mDia2, and human mDia2 (accession numbers O08808, NP_005210, NP_062644, NP_001035982, respectively). Bold lysines represent ubiquitinated residues identified in Denis et al. (29). B, schematic profile of mDia2 mutants examined for ubiquitination. WT, wild type. C, lysine-to-arginine mutant versions of Myc-mDia2 were co-transfected with HA-ubiquitin in HEK293T cells. Lysates were Myc-immunoprecipitated (IP), and immunoblots (IB) were probed with anti-Myc and anti-HA. D, mDia2 mutants were co-transfected with HA-ubiquitin in HEK293T cells. Lysates were immunoprecipitated, and immunoblots were probed with anti-GFP, anti-Myc, or anti-HA to examine the extent of ubiquitination.