ROR and RYK extracellular region structures suggest that receptor tyrosine kinases have distinct WNT-recognition modes
Fumin Shi,
Jeannine M. Mendrola,
Joshua B. Sheetz,
Neo Wu,
Anselm Sommer,
Kelsey F. Speer,
Jasprina N. Noordermeer,
Zhong-Yuan Kan,
Kay Perry,
S. Walter Englander,
Steven E. Stayrook,
Lee G. Fradkin,
Mark A. Lemmon
Affiliations
Fumin Shi
Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA; Graduate Group in Biochemistry and Molecular Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA
Jeannine M. Mendrola
Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA
Joshua B. Sheetz
Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06510, USA; Yale Cancer Biology Institute, Yale University West Campus, West Haven, CT 06516, USA
Neo Wu
Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA; Graduate Group in Biochemistry and Molecular Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA
Anselm Sommer
Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06510, USA; Yale Cancer Biology Institute, Yale University West Campus, West Haven, CT 06516, USA
Kelsey F. Speer
Cell and Molecular Biology Graduate Group, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA; Department of Medicine (Hematology-Oncology), University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA
Jasprina N. Noordermeer
Molecular Cell Biology, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands
Zhong-Yuan Kan
Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA
Kay Perry
NE-CAT, Department of Chemistry and Chemical Biology, Cornell University, Argonne National Laboratory, Argonne, IL 60439, USA
S. Walter Englander
Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA; Graduate Group in Biochemistry and Molecular Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA
Steven E. Stayrook
Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA; Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06510, USA; Yale Cancer Biology Institute, Yale University West Campus, West Haven, CT 06516, USA
Lee G. Fradkin
Molecular Cell Biology, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands; Department of Neurobiology, University of Massachusetts Medical School, Worcester, MA 01655, USA
Mark A. Lemmon
Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA; Graduate Group in Biochemistry and Molecular Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA; Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06510, USA; Yale Cancer Biology Institute, Yale University West Campus, West Haven, CT 06516, USA; Cell and Molecular Biology Graduate Group, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA; Corresponding author
Summary: WNTs play key roles in development and disease, signaling through Frizzled (FZD) seven-pass transmembrane receptors and numerous co-receptors including ROR and RYK family receptor tyrosine kinases (RTKs). We describe crystal structures and WNT-binding characteristics of extracellular regions from the Drosophila ROR and RYK orthologs Nrk (neurospecific receptor tyrosine kinase) and Derailed-2 (Drl-2), which bind WNTs though a FZD-related cysteine-rich domain (CRD) and WNT-inhibitory factor (WIF) domain respectively. Our crystal structures suggest that neither Nrk nor Drl-2 can accommodate the acyl chain typically attached to WNTs. The Nrk CRD contains a deeply buried bound fatty acid, unlikely to be exchangeable. The Drl-2 WIF domain lacks the lipid-binding site seen in WIF-1. We also find that recombinant DWnt-5 can bind Drosophila ROR and RYK orthologs despite lacking an acyl chain. Alongside analyses of WNT/receptor interaction sites, our structures provide further insight into how WNTs may recruit RTK co-receptors into signaling complexes.
