Phosphoprotein phosphatase-2A docks to Dishevelled and counterregulates Wnt3a/β-catenin signaling

Journal of Molecular Signaling - Tập 2 - Trang 1-18 - 2007
Noriko Yokoyama1, Craig C Malbon1
1Department of Pharmacology, Health Sciences Center, State University of New York at Stony Brook, Stony Brook, USA

Tóm tắt

Wnt3a stimulates cellular trafficking of key signaling elements (e.g., Axin, Dishevelled-2, β-catenin, and glycogen synthase kinase-3β) and primitive endoderm formation in mouse F9 embryonic teratocarcinoma cells. The role of phosphoprotein phosphatase-2A in signaling of the Wnt/β-catenin/Lef-Tcf-sensitive gene activation pathway was investigated. Wnt3a action attenuates phosphoprotein phosphatase-2A activity and stimulates the Lef/Tcf-sensitive gene transcription. Inhibiting phosphoprotein phosphatase-2A by okadaic acid, by treatment with siRNA (targeting the C-subunit of the enzyme), or by expression of SV40 small t antigen mimics Wnt3a action, increasing the cellular abundance of Axin and phospho-glycogen synthase kinase-3β as well as the trafficking of signaling elements in the Wnt/β-catenin pathway. Although mimicking effects of Wnt3a on the cellular abundance and trafficking of key signaling elements in the Wnt canonical pathway, suppression of phosphatase-2A alone did not provoke activation of the Lef/Tcf-sensitive transcriptional response, but did potentiate its activation by Wnt3a. Phosphoprotein phosphatase-2A and the scaffold phosphoprotein Dishevelled-2 display similarities in cellular trafficking in response to either Wnt3a or suppression of the phosphatase. A docking site for phosphoprotein phosphatase-2A in the DEP domain of Dishevelled-2 was identified. In current study, we showed new roles of phosphoprotein phosphatase-2A in Wnt/β-catenin signaling pathway: effect on protein expression, effect on protein trafficking, retention of molecules in subcellular compartments, and regulation of enzymatic activity of several key players. Docking of phosphoprotein phosphatase-2A by Dishevelled-2 suppresses phosphatase activity and explains in part the central role of this phosphatase in the counterregulation of the Wnt/β-catenin signaling pathway.

Tài liệu tham khảo

Hunter T: Protein kinases and phosphatases: the yin and yang of protein phosphorylation and signaling. Cell 1995, 80:225–236. Millward TA, Zolnierowicz S, Hemmings BA: Regulation of protein kinase cascades by protein phosphatase 2A. Trend Biochem Sci 1999, 24:186–191. Goldberg Y: Protein phosphatase 2A: who shall regulate the regulator? Biochem Pharmacol 1999, 57:321–328. Wera S, Hemmings BA: Serine/threonine protein phosphatases. Biochem J 1995, 311:17–29. Mayer-Jaekel RE, Hemmings BA: Protein phosphatase 2A-a 'menage a trois'. Trends Cell Biol 1994, 4:287–291. Hsu W, Zeng L, Costantini F: Identification of a domain of Axin that binds to the serine/threonine protein phosphatase 2A and a self-binding domain. J Biol Chem 1999, 274:3439–3445. Ratcliffe MJ, Itoh K, Sokol SY: A positive role for the PP2A catalytic subunit in Wnt signal transduction. J Biol Chem 2000, 275:35680–35683. Willert K, Shibamoto S, Nusse R: Wnt-induced dephosphorylation of axin releases beta-catenin from the axin complex. Genes Dev 1999, 13:1768–1773. Ikeda S, Kishida M, Matsuura Y, Usui H, Kikuchi A: GSK-3beta-dependent phosphorylation of adenomatous polyposis coli gene product can be modulated by beta-catenin and protein phosphatase 2A complexed with Axin. Oncogene 2000, 19:537–545. Sutherland C, Cohen P: The alpha-isoform of glycogen synthase kinase-3 from rabbit skeletal muscle is inactivated by p70 S6 kinase or MAP kinase-activated protein kinase-1 in vitro. FEBS Lett 1994, 338:37–42. Seeling JM, Miller JR, Gil R, Moon RT, White R, Virshup DM: Regulation of beta-catenin signaling by the B56 subunit of protein phosphatase 2A. Science 1999, 283:2089–2091. Li X, Yost HJ, Virshup DM, Seeling JM: Protein phosphatase 2A and its B56 regulatory subunit inhibit Wnt signaling in Xenopus. Embo J 2001, 20:4122–4131. Yang J, Wu J, Tan C, Klein PS: PP2A:B56epsilon is required for Wnt/beta-catenin signaling during embryonic development. Development 2003, 130:5569–5578. Wang J, Wynshaw-Boris A: The canonical Wnt pathway in early mammalian embryogenesis and stem cell maintenance/differentiation. Curr Opin Genet Dev 2004, 14:533–539. Malbon CC: G proteins in development. Nat Rev Mol Cell Biol 2005, 6:689–701. Brantjes H, Barker N, van Es J, Clevers H: TCF: Lady Justice casting the final verdict on the outcome of Wnt signalling. Biol Chem 2002, 383:255–261. Gordon MD, Nusse R: Wnt signaling: multiple pathways, multiple receptors, and multiple transcription factors. J Biol Chem 2006, 281:22429–22433. Wallingford JB, Habas R: The developmental biology of Dishevelled: an enigmatic protein governing cell fate and cell polarity. Development 2005, 132:4421–4436. Liu T, DeCostanzo AJ, Liu X, Wang H, Hallagan S, Moon RT, Malbon CC: G protein signaling from activated rat frizzled-1 to the beta-catenin-Lef-Tcf pathway. Science 2001, 292:1718–1722. Yokoyama N, Dezhong Y, Malbon CC: Abundance, complexation, and trafficking of Wnt/β-catenin signaling elements inresponse to Wnt3a. J Mol Signal 2007.,2(11): Liu T, Liu X, Wang H, Moon RT, Malbon CC: Activation of rat frizzled-1 promotes Wnt signaling and differentiation of mouse F9 teratocarcinoma cells via pathways that require Galpha (q) and Galpha (o) function. J Biol Chem 1999, 274:33539–33544. Li H, Malbon CC, Wang HY: Gene profiling of Frizzled-1 and Frizzled-2 signaling: expression of G-protein-coupled receptor chimeras in mouse F9 teratocarcinoma embryonal cells. Mol Pharmacol 2004, 65:45–55. Yang SI, Lickteig RL, Estes R, Rundell K, Walter G, Mumby MC: Control of protein phosphatase 2A by simian virus 40 small-t antigen. Mol Cell Biol 1991, 11:1988–1995. Mateer SC, Fedorov SA, Mumby MC: Identification of structural elements involved in the interaction of simian virus 40 small tumor antigen with protein phosphatase 2A. J Biol Chem 1998, 273:35339–35346. Cross DA, Alessi DR, Cohen P, Andjelkovich M, Hemmings BA: Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B. Nature 1995, 378:785–789. Frame S, Cohen P, Biondi RM: A common phosphate binding site explains the unique substrate specificity of GSK3 and its inactivation by phosphorylation. Mol Cell 2001, 7:1321–1327. Penton A, Wodarz A, Nusse R: A mutational analysis of dishevelled in Drosophila defines novel domains in the dishevelled protein as well as novel suppressing alleles of axin. Genetics 2002, 161:747–762. Yanagawa S, van Leeuwen F, Wodarz A, Klingensmith J, Nusse R: The dishevelled protein is modified by wingless signaling in Drosophila. Genes Dev 1995, 9:1087–1097. Pallas DC, Shahrik LK, Martin BL, Jaspers S, Miller TB, Brautigan DL, Roberts TM: Polyoma small and middle T antigens and SV40 small t antigen form stable complexes with protein phosphatase 2A. Cell 1990, 60:167–176. Wong HC, Mao J, Nguyen JT, Srinivas S, Zhang W, Liu B, Li L, Wu D, Zheng J: Structural basis of the recognition of the dishevelled DEP domain in the Wnt signaling pathway. Nat Struct Biol 2000, 7:1178–1184. Pan WJ, Pang SZ, Huang T, Guo HY, Wu D, Li L: Characterization of function of three domains in dishevelled-1: DEP domain is responsible for membrane translocation of dishevelled-1. Cell Res 2004, 14:324–330. Schwarz-Romond T, Metcalfe C, Bienz M: Dynamic recruitment of axin by Dishevelled protein assemblies. J Cell Sci 2007, 120:2402–2412. Seto ES, Bellen HJ: Internalization is required for proper Wingless signaling in Drosophila melanogaster. J Cell Biol 2006, 173:95–106. Yokoyama N, Reich NC, Miller WT: Involvement of protein phosphatase 2A in the interleukin-3-stimulated Jak2-Stat5 signaling pathway. J Interferon Cytokine Res 2001, 21:369–378. Xu B, Miller WT: Src homology domains of v-Src stabilize an active conformation of the tyrosine kinase catalytic domain. Mol Cell Biochem 1996, 158:57–63. Chung H, Nairn AC, Murata K, Brautigan DL: Mutation of Tyr307 and Leu309 in the protein phosphatase 2A catalytic subunit favors association with the alpha 4 subunit which promotes dephosphorylation of elongation factor-2. Biochemistry 1999, 38:10371–10376.