Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.


Logo for

J. Biol. Chem. 282 (21): 15903-15911

© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

R-spondin1 Is a High Affinity Ligand for LRP6 and Induces LRP6 Phosphorylation and beta-Catenin Signaling*

Qiou Wei{ddagger}1, Chika Yokota{ddagger}, Mikhail V. Semenov{ddagger}, Brad Doble§, Jim Woodgett§, , and Xi He{ddagger}2

{ddagger}Program of Neurobiology, Children's Hospital Boston, Department of Neurology, Harvard Medical School, Boston, Massachusetts 02115 and the §Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario M5G 1X5, Canada

Abstract: R-spondin proteins are newly identified secreted molecules that activate beta-catenin signaling. However, the mechanism of R-spondin action and its relationship with Wnt signaling remain unclear. Here we show that human R-spondin1 (hRspo1) is a high affinity ligand for the Wnt co-receptor LRP6 (Kd = 1.2 nM). hRspo1 induces glycogen synthase kinase 3-dependent phosphorylation and activation of LRP6. DKK1, an LRP6 antagonist, inhibits hRspo1-induced LRP6 phosphorylation. We further demonstrate that hRspo1 synergizes with Frizzled5 in Xenopus axis induction assays and induces the phosphorylation of Dishevelled, a cytoplasmic component downstream of Frizzled function. Our study reveals interesting similarity and distinction between Wnt and R-spondin signaling.

Received for publication October 10, 2006. Revision received March 30, 2007.

* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1 Postdoctoral Fellow of the Leukemia and Lymphoma Society.

2 Supported by grants from the National Institutes of Health. W. M. Keck Foundation Distinguished Young Scholar and Leukemia and Lymphoma Society Scholar. To whom correspondence should be addressed: Program of Neurobiology, Children's Hospital Boston, Dept. of Neurology, Harvard Medical School, 300 Longwood Ave., Boston, MA 02115. Tel.: 617-919-2257; Fax: 617-730-1953; E-mail: Xi.He{at}

RSPO-LGR4 functions via IQGAP1 to potentiate Wnt signaling.
K. S. Carmon, X. Gong, J. Yi, A. Thomas, and Q. Liu (2014)
PNAS 111, E1221-E1229
   Abstract »    Full Text »    PDF »
Regulation of the Follistatin Gene by RSPO-LGR4 Signaling via Activation of the WNT/{beta}-Catenin Pathway in Skeletal Myogenesis.
X. H. Han, Y.-R. Jin, L. Tan, T. Kosciuk, J.-S. Lee, and J. K. Yoon (2014)
Mol. Cell. Biol. 34, 752-764
   Abstract »    Full Text »    PDF »
The R-spondin/Lgr5/Rnf43 module: regulator of Wnt signal strength.
W. de Lau, W. C. Peng, P. Gros, and H. Clevers (2014)
Genes & Dev. 28, 305-316
   Abstract »    Full Text »    PDF »
Interaction with both ZNRF3 and LGR4 is required for the signalling activity of R-spondin.
Y. Xie, R. Zamponi, O. Charlat, M. Ramones, S. Swalley, X. Jiang, D. Rivera, W. Tschantz, B. Lu, L. Quinn, et al. (2013)
EMBO Rep. 14, 1120-1126
   Abstract »    Full Text »    PDF »
The structural basis of R-spondin recognition by LGR5 and RNF43.
P.-H. Chen, X. Chen, Z. Lin, D. Fang, and X. He (2013)
Genes & Dev. 27, 1345-1350
   Abstract »    Full Text »    PDF »
Tumor-Initiating Cells and FZD8 Play a Major Role in Drug Resistance in Triple-Negative Breast Cancer.
S. Yin, L. Xu, R. D. Bonfil, S. Banerjee, F. H. Sarkar, S. Sethi, and K. B. Reddy (2013)
Mol. Cancer Ther. 12, 491-498
   Abstract »    Full Text »    PDF »
Secreted and Transmembrane Wnt Inhibitors and Activators.
C.-M. Cruciat and C. Niehrs (2013)
Cold Spring Harb Perspect Biol 5, a015081
   Abstract »    Full Text »    PDF »
LGR5 Interacts and Cointernalizes with Wnt Receptors To Modulate Wnt/{beta}-Catenin Signaling.
K. S. Carmon, Q. Lin, X. Gong, A. Thomas, and Q. Liu (2012)
Mol. Cell. Biol. 32, 2054-2064
   Abstract »    Full Text »    PDF »
Both LRP5 and LRP6 Receptors Are Required to Respond to Physiological Wnt Ligands in Mammary Epithelial Cells and Fibroblasts.
S. Goel, E. N. Chin, S. A. Fakhraldeen, S. M. Berry, D. J. Beebe, and C. M. Alexander (2012)
J. Biol. Chem. 287, 16454-16466
   Abstract »    Full Text »    PDF »
NLK positively regulates Wnt/{beta}-catenin signalling by phosphorylating LEF1 in neural progenitor cells.
S. Ota, S. Ishitani, N. Shimizu, K. Matsumoto, M. Itoh, and T. Ishitani (2012)
EMBO J. 31, 1904-1915
   Abstract »    Full Text »    PDF »
Rspo1/Wnt signaling promotes angiogenesis via Vegfc/Vegfr3.
A. V. Gore, M. R. Swift, Y. R. Cha, B. Lo, M. C. McKinney, W. Li, D. Castranova, A. Davis, Y.-s. Mukouyama, and B. M. Weinstein (2011)
Development 138, 4875-4886
   Abstract »    Full Text »    PDF »
LGR4 and LGR5 are R-spondin receptors mediating Wnt/{beta}-catenin and Wnt/PCP signalling.
A. Glinka, C. Dolde, N. Kirsch, Y.-L. Huang, O. Kazanskaya, D. Ingelfinger, M. Boutros, C.-M. Cruciat, and C. Niehrs (2011)
EMBO Rep. 12, 1055-1061
   Abstract »    Full Text »    PDF »
The Regulation of Valvular and Vascular Sclerosis by Osteogenic Morphogens.
K. I. Bostrom, N. M. Rajamannan, and D. A. Towler (2011)
Circ. Res. 109, 564-577
   Abstract »    Full Text »    PDF »
R-spondins function as ligands of the orphan receptors LGR4 and LGR5 to regulate Wnt/{beta}-catenin signaling.
K. S. Carmon, X. Gong, Q. Lin, A. Thomas, and Q. Liu (2011)
PNAS 108, 11452-11457
   Abstract »    Full Text »    PDF »
A WNT/{beta}-Catenin Signaling Activator, R-spondin, Plays Positive Regulatory Roles during Skeletal Myogenesis.
X. H. Han, Y.-R. Jin, M. Seto, and J. K. Yoon (2011)
J. Biol. Chem. 286, 10649-10659
   Abstract »    Full Text »    PDF »
WNT pathways and upper limb anomalies.
M. M. Al-Qattan (2011)
J Hand Surg Eur Vol. 36, 9-22
   Abstract »    Full Text »    PDF »
International Union of Basic and Clinical Pharmacology. LXXX. The Class Frizzled Receptors.
G. Schulte (2010)
Pharmacol. Rev. 62, 632-667
   Abstract »    Full Text »    PDF »
Anterior neural development requires Del1, a matrix-associated protein that attenuates canonical Wnt signaling via the Ror2 pathway.
A. Takai, H. Inomata, A. Arakawa, R. Yakura, M. Matsuo-Takasaki, and Y. Sasai (2010)
Development 137, 3293-3302
   Abstract »    Full Text »    PDF »
R-spondin-1 Is a Novel {beta}-Cell Growth Factor and Insulin Secretagogue.
V. S. C. Wong, A. Yeung, W. Schultz, and P. L. Brubaker (2010)
J. Biol. Chem. 285, 21292-21302
   Abstract »    Full Text »    PDF »
G Protein-coupled Receptor Kinases Phosphorylate LRP6 in the Wnt Pathway.
M. Chen, M. Philipp, J. Wang, R. T. Premont, T. R. Garrison, M. G. Caron, R. J. Lefkowitz, and W. Chen (2009)
J. Biol. Chem. 284, 35040-35048
   Abstract »    Full Text »    PDF »
Wnt11 Promotes Osteoblast Maturation and Mineralization through R-spondin 2.
M. S. Friedman, S. M. Oyserman, and K. D. Hankenson (2009)
J. Biol. Chem. 284, 14117-14125
   Abstract »    Full Text »    PDF »
R-Spondin1 protects mice from chemotherapy or radiation-induced oral mucositis through the canonical Wnt/{beta}-catenin pathway.
J. Zhao, K.-A. Kim, J. De Vera, S. Palencia, M. Wagle, and A. Abo (2009)
PNAS 106, 2331-2336
   Abstract »    Full Text »    PDF »
The Wnt signaling regulator R-spondin 3 promotes angioblast and vascular development.
O. Kazanskaya, B. Ohkawara, M. Heroult, W. Wu, N. Maltry, H. G. Augustin, and C. Niehrs (2008)
Development 135, 3655-3664
   Abstract »    Full Text »    PDF »
Differential Regulation of STAT Family Members by Glycogen Synthase Kinase-3.
E. Beurel and R. S. Jope (2008)
J. Biol. Chem. 283, 21934-21944
   Abstract »    Full Text »    PDF »
Targeted Disruption of the Wnt Regulator Kremen Induces Limb Defects and High Bone Density.
K. Ellwanger, H. Saito, P. Clement-Lacroix, N. Maltry, J. Niedermeyer, W. K. Lee, R. Baron, G. Rawadi, H. Westphal, and C. Niehrs (2008)
Mol. Cell. Biol. 28, 4875-4882
   Abstract »    Full Text »    PDF »
Crosstalk between AHR and Wnt signaling through R-Spondin1 impairs tissue regeneration in zebrafish.
L. K. Mathew, S. S. Sengupta, J. LaDu, E. A. Andreasen, and R. L. Tanguay (2008)
FASEB J 22, 3087-3096
   Abstract »    Full Text »    PDF »
DKK1 Antagonizes Wnt Signaling without Promotion of LRP6 Internalization and Degradation.
M. V. Semenov, X. Zhang, and X. He (2008)
J. Biol. Chem. 283, 21427-21432
   Abstract »    Full Text »    PDF »
Wnt Signal Amplification via Activity, Cooperativity, and Regulation of Multiple Intracellular PPPSP Motifs in the Wnt Co-receptor LRP6.
B. T. MacDonald, C. Yokota, K. Tamai, X. Zeng, and X. He (2008)
J. Biol. Chem. 283, 16115-16123
   Abstract »    Full Text »    PDF »
R-Spondin Family Members Regulate the Wnt Pathway by a Common Mechanism.
K.-A. Kim, M. Wagle, K. Tran, X. Zhan, M. A. Dixon, S. Liu, D. Gros, W. Korver, S. Yonkovich, N. Tomasevic, et al. (2008)
Mol. Biol. Cell 19, 2588-2596
   Abstract »    Full Text »    PDF »
Activation of {beta}-catenin signaling by Rspo1 controls differentiation of the mammalian ovary.
A.-A. Chassot, F. Ranc, E. P. Gregoire, H. L. Roepers-Gajadien, M. M. Taketo, G. Camerino, D. G. de Rooij, A. Schedl, and M.-C. Chaboissier (2008)
Hum. Mol. Genet. 17, 1264-1277
   Abstract »    Full Text »    PDF »
R-spondin1 plays an essential role in ovarian development through positively regulating Wnt-4 signaling.
K. Tomizuka, K. Horikoshi, R. Kitada, Y. Sugawara, Y. Iba, A. Kojima, A. Yoshitome, K. Yamawaki, M. Amagai, A. Inoue, et al. (2008)
Hum. Mol. Genet. 17, 1278-1291
   Abstract »    Full Text »    PDF »
R-spondin 2 is required for normal laryngeal-tracheal, lung and limb morphogenesis.
S. M. Bell, C. M. Schreiner, S. E. Wert, M. L. Mucenski, W. J. Scott, and J. A. Whitsett (2008)
Development 135, 1049-1058
   Abstract »    Full Text »    PDF »
Initiation of Wnt signaling: control of Wnt coreceptor Lrp6 phosphorylation/activation via frizzled, dishevelled and axin functions.
X. Zeng, H. Huang, K. Tamai, X. Zhang, Y. Harada, C. Yokota, K. Almeida, J. Wang, B. Doble, J. Woodgett, et al. (2008)
Development 135, 367-375
   Abstract »    Full Text »    PDF »
Analysis of Endogenous LRP6 Function Reveals a Novel Feedback Mechanism by Which Wnt Negatively Regulates Its Receptor.
Z. Khan, S. Vijayakumar, T. V. de la Torre, S. Rotolo, and A. Bafico (2007)
Mol. Cell. Biol. 27, 7291-7301
   Abstract »    Full Text »    PDF »
WNTers in La Jolla.
S. Y. Sokol and K. A. Wharton Jr (2007)
Development 134, 3393-3399
   Abstract »    Full Text »    PDF »
R-Spondin1 regulates Wnt signaling by inhibiting internalization of LRP6.
M. E. Binnerts, K.-A. Kim, J. M. Bright, S. M. Patel, K. Tran, M. Zhou, J. M. Leung, Y. Liu, W. E. Lomas III, M. Dixon, et al. (2007)
PNAS 104, 14700-14705
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882