title: ADAM17-mediated shedding of the IL6R induces cleavage of the membrane stub by gamma-secretase.
abstract: Interleukin-6 (IL6) signals are mediated by classic and trans-signaling. In classic signaling, IL6 first binds to the membrane bound Interleukin-6 Receptor (IL6R) whereas in trans-signaling, IL6 acts via a soluble form of the IL6R. Trans-signaling via the soluble IL6R (sIL6R) was linked to chronic inflammation and cancer. The release of the IL6R is mediated by the disintegrin and metalloproteinases ADAM10 and ADAM17. To analyze the fate of the C-terminal cleavage fragment after ectodomain shedding we fused the IL6R C-terminally to two Z-domains of Protein-A (2Z-tag) or to GFP. A specific C-terminal fragment of the IL6R protein could be detected after ADAM17-induced shedding. Using gamma-secretase inhibitors and gene-deficient cells, we demonstrate that after ADAM17 mediated cleavage, the IL6R C-terminal fragment was cleaved by the gamma-secretase at the plasma membrane. We were, however, not able to detect an IL6R intracellular domain. After gamma-secretase cleavage IL6R cell surface expression was lost and gamma-secretase cleavage product(s) of the IL6R were endocytosed. No GFP-fluorescence of a gamma-secretase-cleaved IL6R-GFP fusion protein was observed in the nucleus. We therefore hypothesize that a potential IL6R intracellular domain fragment is not involved in nuclear signaling but rapidly degraded.
authors.affiliation: Institute of Biochemistry, Christian-Albrechts-University of Kiel, Olshausenstr. 40, D-24098 Kiel, Germany.
authors.forename: Athena
authors.initials: A
authors.lastname: Chalaris
authors.forename: Jessica
authors.initials: J
authors.lastname: Gewiese
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Paliga
authors.forename: Lina
authors.initials: L
authors.lastname: Fleig
authors.forename: Alex
authors.initials: A
authors.lastname: Schneede
authors.forename: Karsten
authors.initials: K
authors.lastname: Krieger
authors.forename: Stefan
authors.initials: S
authors.lastname: Rose-John
authors.forename: Jürgen
authors.initials: J
authors.lastname: Scheller
mesh_terms: D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism*; D000072198:ADAM17 Protein; D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D005231:Fatty Acids, Unsaturated / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism; D019947:Receptors, Interleukin-6 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology
chemical_list: D005231:Fatty Acids, Unsaturated; D007527:Isoenzymes; D010446:Peptide Fragments; D019947:Receptors, Interleukin-6; D011993:Recombinant Fusion Proteins; C038752:leptomycin A; D011493:Protein Kinase C; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072198:ADAM17 Protein; C000606462:ADAM17 protein, human