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Protein Interaction Profiling of the p97 Adaptor UBXD1 Points to a Role for the Complex in Modulating ERGIC-53 Trafficking*
| Content Provider | PubMed Central |
|---|---|
| Author | Haines, Dale S. Lee, J. Eugene Beauparlant, Stephen L. Kyle, Dane B. Besten, Willem Den Sweredoski, Michael J. Graham, Robert L. J. Hess, Sonja Deshaies, Raymond J. |
| Copyright Year | 2012 |
| Abstract | UBXD1 is a member of the poorly understood subfamily of p97 adaptors that do not harbor a ubiquitin association domain or bind ubiquitin-modified proteins. Of clinical importance, p97 mutants found in familial neurodegenerative conditions Inclusion Body Myopathy Paget's disease of the bone and/or Frontotemporal Dementia and Amyotrophic Lateral Sclerosis are defective at interacting with UBXD1, indicating that functions regulated by a p97-UBXD1 complex are altered in these diseases. We have performed liquid chromatography-mass spectrometric analysis of UBXD1-interacting proteins to identify pathways in which UBXD1 functions. UBXD1 displays prominent association with ERGIC-53, a hexameric type I integral membrane protein that functions in protein trafficking. The UBXD1-ERGIC-53 interaction requires the N-terminal 10 residues of UBXD1 and the C-terminal cytoplasmic 12 amino acid tail of ERGIC-53. Use of p97 and E1 enzyme inhibitors indicate that complex formation between UBXD1 and ERGIC-53 requires the ATPase activity of p97, but not ubiquitin modification. We also performed SILAC-based quantitative proteomic profiling to identify ERGIC-53 interacting proteins. This analysis identified known (e.g. COPI subunits) and novel (Rab3GAP1/2 complex involved in the fusion of vesicles at the cell membrane) interactions that are also mediated through the C terminus of the protein. Immunoprecipitation and Western blotting analysis confirmed the proteomic interaction data and it also revealed that an UBXD1-Rab3GAP association requires the ERGIC-53 binding domain of UBXD1. Localization studies indicate that UBXD1 modules the sub-cellular trafficking of ERGIC-53, including promoting movement to the cell membrane. We propose that p97-UBXD1 modulates the trafficking of ERGIC-53-containing vesicles by controlling the interaction of transport factors with the cytoplasmic tail of ERGIC-53. |
| Related Links | http://dx.doi.org/10.1074/mcp.m111.016444 |
| File Format | |
| ISSN | 15359476 |
| e-ISSN | 15359484 |
| Journal | Molecular & Cellular Proteomics : MCP |
| Issue Number | 6 |
| Volume Number | 11 |
| Language | English |
| Publisher | The American Society for Biochemistry and Molecular Biology |
| Publisher Date | 2012-06-01 |
| Access Restriction | Open |
| Rights Holder | The American Society for Biochemistry and Molecular Biology |
| Subject Keyword | Analytical Chemistry Biochemistry Molecular Biology Research in Higher Education |
| Content Type | Text |
| Resource Type | Article |
| Subject | Medicine Analytical Chemistry Molecular Biology Biochemistry |
