NIN1 is an essential gene for growth of the yeast Saccharomyces cerevisiae and was recently found to encode a component of the regulatory subunit of the 26S proteasome. The nin1-1 mutant is temperature sensitive and its main defect is in G1/S progression and G2/M progression at non-permissive temperatures. One of the two multicopy suppressors of nin1-1, SUN2 (SUppressor of Nin1-1), was found to encode a protein of 523 amino acids whose sequence is similar to those of Drosophila melanogaster diphenol oxidase A2 and the mouse mast-cell Tum(-) transplantation antigen, P91A. The C-terminal half of Sun2p was found to be functional as Sun2p at 25 degrees C, 30 degrees C, and 34 degrees C but not at 37 degrees C. The open reading frame (ORF) of the Drosophila diphenol oxidase A2 gene (Dox-A2) was obtained from a lambda phage cDNA library using the polymerase chain reaction technique. The Dox-A2 ORF driven by the TDH3 promoter complemented the phenotype of a strain deleted for sun2. This Dox-A2-dependent strain was temperature sensitive and accumulated dumb-bell-shaped cells, with an undivided nucleus at the isthmus, after temperature upshift. This morphology is similar to that of nin1-1 cells kept at a restrictive temperature. These results suggest that SUN2 is a functional counterpart of Dox-A2 and that these genes play a pivotal role in the cell cycle in each organism.