A novel centrosomal RING-finger protein, Dorfin, mediates ubiquitin ligase activity

We cloned a novel gene, Dorfin (double ring-finger protein), from human spinal cord. The Dorfin mRNA transcript was 4.4 kb and expressed ubiquitously in many organs as well as in the central nervous system, including the spinal cord. Dorfin encoded 838 amino acid protein Dorfin, which contains two RING-finger motifs and an IBR (in between RING-fingers) motif at its N-terminus. Dorfin is a short-lived protein. Treatment with MG132, a potent proteasome inhibitor, resulted in the accumulation of ubiquitinated Dorfin and Dorfin-associated cellular proteins in cultured cells. Dorfin bound specifically with human ubiquitin-conjugating enzymes UbcH7 and UbcH8 through the RING-finger/IBR domain. Partial deletion of the RING-finger/IBR domain eliminated these interaction and ubiquitination activities. These results strongly suggest that Dorfin is a new member of RING-finger type ubiquitin ligase. Dorfin is localized in the centrosome and probably functions in the microtubule organizing centers. (C) 2001 Academic Press.