Targeted truncation of the ADAM17 cytoplasmic domain in mice results in protein destabilization and a hypomorphic phenotype Journal Article


Authors: Lora, J.; Weskamp, G.; Li, T. M.; Maretzky, T.; Shola, D. T. N.; Monette, S.; Lichtenthaler, S. F.; Lu, T. T.; Yang, C.; Blobel, C. P.
Article Title: Targeted truncation of the ADAM17 cytoplasmic domain in mice results in protein destabilization and a hypomorphic phenotype
Abstract: 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. © 2021 THE AUTHORS.
Keywords: cell proliferation; proteins; cell death; tumor necrosis factor alpha; cell culture; bone; cell membranes; macrophages; mammals; signaling pathways; cell signaling; mouse embryonic fibroblasts; activation analysis; epidermal growth factor receptors; trans-membrane domains; cytoplasmic domains; functional assays; protein destabilization
Journal Title: Journal of Biological Chemistry
Volume: 296
ISSN: 0021-9258
Publisher: American Society for Biochemistry and Molecular Biology  
Date Published: 2021-01-01
Start Page: 100733
Language: English
DOI: 10.1016/j.jbc.2021.100733
PUBMED: 33957124
PROVIDER: scopus
PMCID: PMC8191336
DOI/URL:
Notes: Article -- Export Date: 1 July 2021 -- Source: Scopus
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  1. Sebastien Monette
    149 Monette