Activity of MDM2, a ubiquitin ligase, toward p53 or itself is dependent on the RING finger domain of the ligase

R Honda, H Yasuda - Oncogene, 2000 - nature.com
R Honda, H Yasuda
Oncogene, 2000nature.com
We previously showed that oncoprotein MDM2 has ubiquitin ligase activity toward tumor
suppressor p53. In that paper, we showed very weak homology in the carboxyl terminal
portion between MDM2 and E6AP (HECT domain). We mutated the cysteine residue (C464)
corresponding to the residue essential for the ubiquitin ligase activity of E6AP and this
mutation diminished the ligase activity of MDM2. The cysteine residue described above is
also one of the cysteine residues that form the RING finger domain of MDM2. We tried to find …
Abstract
We previously showed that oncoprotein MDM2 has ubiquitin ligase activity toward tumor suppressor p53. In that paper, we showed very weak homology in the carboxyl terminal portion between MDM2 and E6AP (HECT domain). We mutated the cysteine residue (C464) corresponding to the residue essential for the ubiquitin ligase activity of E6AP and this mutation diminished the ligase activity of MDM2. The cysteine residue described above is also one of the cysteine residues that form the RING finger domain of MDM2. We tried to find out whether the diminishing of the activity by the mutation is attributable to the disruption of the RING finger domain or not. When the ring finger domain of MDM2 was deleted, the truncation mutant did not have the ubiquitin ligase activity. When we mutated the seven cysteine residues of RING finger domain of MDM2 in the carboxyl terminus, the disruption of each residue in the RING finger completely diminished the ubiquitin ligase activity of MDM2 toward MDM2 itself and toward tumor suppressor p53. These data indicate that the RING finger domain in MDM2 is essential for its ubiquitin ligase activity toward p53 and itself.
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