Targeted truncation of the ADAM17 cytoplasmic domain in mice results in protein destabilization and a hypomorphic phenotype
| العنوان: | Targeted truncation of the ADAM17 cytoplasmic domain in mice results in protein destabilization and a hypomorphic phenotype |
|---|---|
| المؤلفون: | Thorsten Maretzky, Stefan F. Lichtenthaler, Gisela Weskamp, Theresa T. Lu, Sebastien Monette, Jose Lora, Dorjee T.N. Shola, Chingwen Yang, Carl P. Blobel, Thomas M. Li |
| المصدر: | The Journal of Biological Chemistry The journal of biological chemistry 296, 100733-(2021). doi:10.1016/j.jbc.2021.100733 |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0301 basic medicine, Male, Cytoplasm, KI, knock-in, knock-in mutation in mice, Biochemistry, Mice, iRhom1 and 2, inactive Rhomboid proteins 1 and 2, ADAM17 Protein, membrane protein, myeloid cell, MFI, mean fluorescence intensity, Receptor, Sequence Deletion, TACE (TNFα converting enzyme), Bone growth, mEF, mouse embryonic fibroblast, Chemistry, Protein Stability, ddc, Cell biology, BMDM, bone-marrow-derived macrophage, TNFα, tumor necrosis factor α, Transmembrane domain, Phenotype, Protein destabilization, ddc:540, DV, donor vector, Female, Signal transduction, metabolism [Fibroblasts], Research Article, IL-6R, interleukin-6 receptor, PMA, phorbol 12-myristate 13-acetate, genetics [ADAM17 Protein], ADAM17 (a disintegrin and metalloprotease 17), 03 medical and health sciences, metabolism [ADAM17 Protein], cell surface enzyme, Protein Domains, CHX, cycloheximide, Gene knockin, Animals, Amino Acid Sequence, Molecular Biology, metabolism [Cytoplasm], 030102 biochemistry & molecular biology, Base Sequence, Cell Biology, chemistry [ADAM17 Protein], Sheddase, Fibroblasts, EGFR, epidermal growth factor receptor, Mice, Inbred C57BL, 030104 developmental biology, EPCAM, epithelial cell adhesion molecule, mES, mouse embryonic stem, CRISPR-Cas Systems, ADAM17, a disintegrin and metalloprotease 17 |
| الوصف: | A disintegrin and metalloprotease 17 (ADAM17) is a cell-surface metalloprotease that serves as the principle sheddase for tumor necrosis factor α (TNFα), interleukin-6 receptor (IL-6R), and several ligands of the epidermal growth factor receptor (EGFR), regulating these crucial signaling pathways. ADAM17 activation requires its transmembrane domain, but not its cytoplasmic domain, and little is known about the role of this domain in vivo. To investigate, we used CRISPR-Cas9 to mutate the endogenous Adam17 locus in mice to produce a mutant ADAM17 lacking its cytoplasmic domain (Adam17Δcyto). Homozygous Adam17Δcyto animals were born at a Mendelian ratio and survived into adulthood with slightly wavy hair and curled whiskers, consistent with defects in ADAM17/EGFR signaling. At birth, Adam17Δcyto mice resembled Adam17−/− mice in that they had open eyes and enlarged semilunar heart valves, but they did not have bone growth plate defects. The deletion of the cytoplasmic domain resulted in strongly decreased ADAM17 protein levels in all tissues and cells examined, providing a likely cause for the hypomorphic phenotype. In functional assays, Adam17Δcyto mouse embryonic fibroblasts and bone-marrow-derived macrophages had strongly reduced ADAM17 activity, consistent with the reduced protein levels. Nevertheless, ADAM17Δcyto could be stimulated by PMA, a well-characterized posttranslational activator of ADAM17, corroborating that the cytoplasmic domain of endogenous ADAM17 is not required for its rapid response to PMA. Taken together, these results provide the first evidence that the cytoplasmic domain of ADAM17 plays a pivotal role in vivo in regulating ADAM17 levels and function. |
| وصف الملف: | application/pdf |
| تدمد: | 1083-351X |
| DOI: | 10.1016/j.jbc.2021.100733 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::48ddba9cdef79f06d77f3f2a019dddab https://pubmed.ncbi.nlm.nih.gov/33957124 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....48ddba9cdef79f06d77f3f2a019dddab |
| قاعدة البيانات: | OpenAIRE |
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