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Identification of developmentally expressed proteins that functionally interact with Nedd4 ubiquitin ligase.
- Source :
-
The Journal of biological chemistry [J Biol Chem] 2002 Jan 25; Vol. 277 (4), pp. 2897-907. Date of Electronic Publication: 2001 Nov 20. - Publication Year :
- 2002
-
Abstract
- Nedd4 is a HECT domain-containing ubiquitin ligase that mediates ubiquitylation and proteasome degradation of target proteins. The molecular basis for the interaction of Nedd4 with substrates lies in its WW domains, which can bind proline-rich (PY) domains in target proteins. Nedd4 is a developmentally expressed protein and may have a fundamental role to play in embryonic processes. However, whether Nedd4 has such a function is currently unknown, in part because few developmentally regulated ubiquitylation substrates have been identified or characterized. We have carried out a yeast two-hybrid screen and identified four proteins expressed in the mid-gestation embryo that are able to interact with Nedd4. Characterization of their functional interaction with Nedd4 in vitro and in vivo demonstrated that three of the four are bona fide Nedd4 binding partners, and two have the capacity to be ubiquitylation substrates. One of these is the first identified nonviral substrate for Nedd4-mediated monoubiquitylation. Interestingly, neither of these two ubiquitylated proteins interacts with Nedd4 through PY-mediated mechanisms. For one of the three Nedd4 binding partners, there was no discernable evidence of ubiquitylation. However, this protein clearly associates with Nedd4 through its PY domains and can alter the location of Nedd4 in cells, suggesting a role other than as a ubiquitylation substrate.
- Subjects :
- Blotting, Western
Cell Line
Cell Nucleus metabolism
Cysteine Endopeptidases
DNA, Complementary metabolism
Endosomal Sorting Complexes Required for Transport
Expressed Sequence Tags
Glutathione Transferase metabolism
Humans
Microscopy, Fluorescence
Multienzyme Complexes antagonists & inhibitors
Mutagenesis, Site-Directed
Nedd4 Ubiquitin Protein Ligases
Plasmids metabolism
Precipitin Tests
Proteasome Endopeptidase Complex
Protein Binding
Protein Biosynthesis
Protein Structure, Tertiary
Recombinant Fusion Proteins metabolism
Transfection
Two-Hybrid System Techniques
Ubiquitin metabolism
Calcium-Binding Proteins metabolism
Ligases metabolism
Ubiquitin-Protein Ligases
Subjects
Details
- Language :
- English
- ISSN :
- 0021-9258
- Volume :
- 277
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- The Journal of biological chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 11717310
- Full Text :
- https://doi.org/10.1074/jbc.M110047200