Structure and function of the ROR2 cysteine-rich domain in vertebrate noncanonical WNT5A signaling

Abstract

The receptor tyrosine kinase ROR2 mediates noncanonical WNT5A signaling to orchestrate tissue morphogenetic processes, and dysfunction of the pathway causes Robinow syndrome, Brachydactyly B and metastatic diseases. The domain(s) and mechanisms required for ROR2 function, however, remain unclear. We solved the crystal structure of the extracellular cysteine-rich (CRD) and Kringle (Kr) domains of ROR2 and found that, unlike other CRDs, the ROR2 CRD lacks the signature hydrophobic pocket that binds lipids/lipid-modified proteins, such as WNTs, suggesting a novel mechanism of ligand reception. Functionally, we showed that the ROR2 CRD, but not other domains, is required and minimally sufficient to promote WNT5A signaling, and Robinow mutations in the CRD and the adjacent Kr impair ROR2 secretion and function. Moreover, using function-activating and -perturbing antibodies against the Frizzled (FZ) family of WNT receptors, we demonstrate the involvement of FZ in WNT5A-ROR signaling. Thus, ROR2 acts via its CRD to potentiate the function of a receptor super-complex that includes FZ to transduce WNT5A signals.

An efficient method for immortalizing mouse embryonic fibroblasts

“…We have successfully used this protocol to generate more than 15 independent iMEF lines, including that described in Griffiths et al, eLife , 2024 (Figures 3 to 5, and the associated supplementary data figures) (Griffiths et al, 2024).…”

“…This study was supported by the National Institutes of Health (1R35GM119574, 1R35GM144341) to H.H.H. This protocol was established to generate the iMEFs used in the following publication: Griffiths et al (2024)…”