title: A transmembrane leucine zipper is required for activation of the dimeric receptor tyrosine kinase DDR1.
abstract: Receptor tyrosine kinases of the discoidin domain family, DDR1 and DDR2, are activated by different types of collagen and play important roles in cell adhesion, migration, proliferation, and matrix remodeling. In a previous study, we found that collagen binding by the discoidin domain receptors (DDRs) requires dimerization of their extracellular domains (Leitinger, B. (2003) J. Biol. Chem. 278, 16761-16769), indicating that the paradigm of ligand-induced receptor dimerization may not apply to the DDRs. Using chemical cross-linking and co-immunoprecipitation of differently tagged DDRs, we now show that the DDRs form ligand-independent dimers in the biosynthetic pathway and on the cell surface. We further show that both the extracellular and the cytoplasmic domains are individually dispensable for DDR1 dimerization. The DDR1 transmembrane domain contains two putative dimerization motifs, a leucine zipper and a GXXXG motif. Mutations disrupting the leucine zipper strongly impaired collagen-induced transmembrane signaling, although the mutant DDR1 proteins were still able to dimerize, whereas mutation of the GXXXG motif had no effect. A bacterial reporter assay (named TOXCAT) showed that the DDR1 transmembrane domain has a strong potential for self-association in a biological membrane and that this interaction occurs via the leucine zipper and not the GXXXG motif. Our results demonstrate that the DDRs exist as stable dimers in the absence of ligand and that receptor activation requires specific interactions made by the transmembrane leucine zipper.
authors.affiliation: Department of Medicine, University College London, 5 University Street, London WC1E 6JJ, UK.
authors.forename: Nafeesa A
authors.initials: NA
authors.lastname: Noordeen
authors.forename: Federico
authors.initials: F
authors.lastname: Carafoli
authors.forename: Erhard
authors.initials: E
authors.lastname: Hohenester
authors.forename: Michael A
authors.initials: MA
authors.lastname: Horton
authors.forename: Birgit
authors.initials: B
authors.lastname: Leitinger
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003094:Collagen / Q000737:chemistry / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D019281:Dimerization; D000070819:Discoidin Domain Receptor 1; D006801:Humans; D007930:Leucine / Q000737:chemistry*; D016350:Leucine Zippers; D008969:Molecular Sequence Data; D009154:Mutation; D051381:Rats; D020794:Receptor Protein-Tyrosine Kinases / Q000737:chemistry*
chemical_list: D003094:Collagen; C092030:DDR1 protein, human; D000070819:Discoidin Domain Receptor 1; D020794:Receptor Protein-Tyrosine Kinases; D007930:Leucine