Cbl interacts with multiple E2s in vitro and in cells
| Autor: | Donna Voeller, Rachel E. Klevit, Ke Ma, Mariya S. Liyasova, Jinqiu Chen, Philip E. Ryan, Stanley Lipkowitz |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Cultured tumor cells
Syk Plasma protein binding environment and public health Biochemistry Receptor tyrosine kinase Ubiquitin hemic and lymphatic diseases Yeast Two-Hybrid Assays Two-Hybrid Screening Monoubiquitination Small interfering RNAs Proto-Oncogene Proteins c-cbl Phosphorylation Post-Translational Modification 0303 health sciences Multidisciplinary biology Chemistry 030302 biochemistry & molecular biology Cell biology Precipitation Techniques ErbB Receptors Nucleic acids Medicine Cell lines biological phenomena cell phenomena and immunity Biological cultures Protein Interaction Assays Proto-oncogene tyrosine-protein kinase Src Protein Binding Research Article Science Immunoblotting Molecular Probe Techniques Library Screening Research and Analysis Methods 03 medical and health sciences Two-Hybrid System Techniques Genetics Humans Immunoprecipitation Gene Silencing HeLa cells Molecular Biology Techniques Non-coding RNA Molecular Biology 030304 developmental biology Molecular Biology Assays and Analysis Techniques fungi Ubiquitination Biology and Life Sciences Proteins Cell cultures Gene regulation enzymes and coenzymes (carbohydrates) HEK293 Cells Gene Expression Regulation Mutation Ubiquitin-Conjugating Enzymes biology.protein RNA UBE2E Family Gene expression |
| Zdroj: | PLoS ONE PLoS ONE, Vol 14, Iss 5, p e0216967 (2019) |
| ISSN: | 1932-6203 |
| Popis: | Many receptor tyrosine kinases (RTKs, such as EGFR, MET) are negatively regulated by ubiquitination and degradation mediated by Cbl proteins, a family of RING finger (RF) ubiquitin ligases (E3s). Loss of Cbl protein function is associated with malignant transformation driven by increased RTK activity. RF E3s, such as the Cbl proteins, interact with a ubiquitin-conjugating enzyme (E2) to confer specificity to the ubiquitination process and direct the transfer of ubiquitin from the E2 to one or more lysines on the target proteins. Using in vitro E3 assays and yeast two-hybrid screens, we found that Ube2d, Ube2e families, Ube2n/2v1, and Ube2w catalyze autoubiquitination of the Cbl protein and Ube2d2, Ube2e1, and Ube 2n/2v1 catalyze Cbl-mediated substrate ubiquitination of the EGFR and SYK. Phosphorylation of the Cbl protein by by Src resulted in increased E3 activity compared to unphosphorylated cbl or Cbl containing a phosphomimetic Y371E mutation. Ubiquitin chain formation depended on the E2 tested with Cbl with Ube2d2 forming both K48 and K63 linked chains, Ube2n/2v1 forming only K63 linked chains, and Ube2w inducing monoubiquitination. In cells, the Ube2d family, Ube2e family, and Ube2n/2v1 contributed to EGFR ubiquitination. Our data suggest that multiple E2s can interact with Cbl and modulate its E3 activity in vitro and in cells. |
| Databáze: | OpenAIRE |
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