Stabilization of the E3 Ubiquitin Ligase Nrdp1 by the Deubiquitinating Enzyme USP8
| Autor: | Xiuli Wu, Kermit L. Carraway, Colleen A Sweeney, Lisa Irwin, Lily Yen |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Ubiquitin-Protein Ligases
Biology Receptor tyrosine kinase Cell Line Deubiquitinating enzyme Mice Affinity chromatography Endopeptidases Enzyme Stability Ring finger medicine Animals Humans ERBB3 Cell Growth and Development Molecular Biology ERBB4 Endosomal Sorting Complexes Required for Transport Ubiquitin Proteins Cell Biology Protein Structure Tertiary Cell biology Ubiquitin ligase RING finger domain medicine.anatomical_structure Biochemistry biology.protein Ubiquitin Thiolesterase Protein Binding |
| Zdroj: | Wu, Xiuli; Yen, Lily; Irwin, Lisa; Sweeney, Colleen; & Carraway, Kermit L. (2004). Stabilization of the E3 ubiquitin ligase Nrdp1 by the deubiquitinating enzyme USP8. Molecular and Cellular Biology, 24(17), 7748-7757. UC Davis: Retrieved from: http://www.escholarship.org/uc/item/3xq0b1j0 |
| ISSN: | 1098-5549 |
| DOI: | 10.1128/mcb.24.17.7748-7757.2004 |
| Popis: | Nrdp1 is a RING finger-containing E3 ubiquitin ligase that physically interacts with and regulates steady-state cellular levels of the ErbB3 and ErbB4 receptor tyrosine kinases and has been implicated in the degradation of the inhibitor-of-apoptosis protein BRUCE. Here we demonstrate that the Nrdp1 protein undergoes efficient proteasome-dependent degradation and that mutations in its RING finger domain that disrupt ubiquitin ligase activity enhance stability. These observations suggest that Nrdp1 self-ubiquitination and stability could play an important role in regulating the activity of this protein. Using affinity chromatography, we identified the deubiquitinating enzyme USP8 (also called Ubpy) as a protein that physically interacts with Nrdp1. Nrdp1 and USP8 could be coimmunoprecipitated, and in transfected cells USP8 specifically bound to Nrdp1 but not cbl, a RING finger E3 ligase involved in ligand-stimulated epidermal growth factor receptor down-regulation. The USP8 rhodanese and catalytic domains mediated Nrdp1 binding. USP8 markedly enhanced the stability of Nrdp1, and a point mutant that disrupts USP8 catalytic activity destabilized endogenous Nrdp1. Our results indicate that Nrdp1 is a specific target for the USP8 deubiquitinating enzyme and are consistent with a model where USP8 augments Nrdp1 activity by mediating its stabilization. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|