The role of the cysteine-rich domain of Frizzled in Wingless-Armadillo signaling.
The Frizzled (Fz) receptors contain seven transmembrane helices and an amino-terminal cysteine-rich domain (CRD) that is sufficient and necessary for binding of the ligands, the Wnts. Recent genetic experiments have suggested, however, that the CRD is dispensable for signaling. We engineered fz CRD mutant transgenes and tested them for Wg signaling activity. None of the mutants was functional in cell culture or could fully replace fz in vivo. We also show that replacing the CRD with a structurally distinct Wnt-binding domain, the Wnt inhibitory factor, reconstitutes a functional Wg receptor. We therefore hypothesized that the function of the CRD is to bring Wg in close proximity with the membrane portion of the receptor. We tested this model by substituting Wg itself for the CRD, a manipulation that results in a constitutively active receptor. We propose that Fz activates signaling in two steps: Fz uses its CRD to capture Wg, and once bound Wg interacts with the membrane portion of the receptor to initiate signaling.
Povelones
Michael
M
Department of Developmental Biology, Beckman Center, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, CA 94305-5428, USA.
Nusse
Roel
R
D000595:Amino Acid Sequence; D000818:Animals; D051186:Armadillo Domain Proteins / Q000378:metabolism; D015153:Blotting, Western; D003545:Cysteine / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers; D004330:Drosophila / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D051157:Frizzled Receptors; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010641:Phenotype; D017434:Protein Structure, Tertiary; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D034622:RNA Interference; D043562:Receptors, G-Protein-Coupled; D015398:Signal Transduction / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism; D019076:Transgenes / Q000235:genetics; D014921:Wings, Animal / Q000033:anatomy & histology; D051155:Wnt1 Protein
C495305:ARM protein, Drosophila; D051186:Armadillo Domain Proteins; D017931:DNA Primers; D029721:Drosophila Proteins; D051157:Frizzled Receptors; D008565:Membrane Proteins; D011518:Proto-Oncogene Proteins; D043562:Receptors, G-Protein-Coupled; D014157:Transcription Factors; D051155:Wnt1 Protein; C058784:fz protein, Drosophila; C074622:wg protein, Drosophila; D003545:Cysteine