Multisite Protein Kinase A and Glycogen Synthase Kinase 3beta Phosphorylation Leads to Gli3 Ubiquitination by SCFbetaTrCP

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 2006 May 18

PMID 16705181

Citations 130

Authors

Denis Tempe

Mariana Casas

Sonia Karaz

Marie-Francoise Blanchet-Tournier

Jean-Paul Concordet

Affiliations

Soon will be listed here.

Abstract

Gli3 is a zinc finger transcription factor proteolytically processed into a truncated repressor lacking C-terminal activation domains. Gli3 processing is stimulated by protein kinase A (PKA) and inhibited by Hedgehog signaling, a major signaling pathway in vertebrate development and disease. We show here that multisite glycogen synthase kinase 3beta (GSK3beta) phosphorylation and ubiquitination by SCFbetaTrCP are required for Gli3 processing. We identified multiple betaTrCP-binding sites related to the DSGX2-4S motif in Gli3, which are intertwined with PKA and GSK3beta sites, and SCFbetaTrCP target lysines that are essential for processing. Our results support a simple model whereby PKA triggers a cascade of Gli3 phosphorylation by GSK3beta and CK1 that leads to direct betaTrCP binding and ubiquitination by SCFbetaTrCP. Binding of betaTrCP to Gli3 N- and C-terminal domains lacking DSGX2-4S-related motifs was also observed, which could reflect indirect interaction via other components of Hedgehog signaling, such as the tumor suppressor Sufu. Gli3 therefore joins a small set of transcription factors whose processing is regulated by the ubiquitin-proteasome pathway. Our study sheds light on the role of PKA phosphorylation in Gli3 processing and will help to analyze how dose-dependent tuning of Gli3 processing is achieved by Hedgehog signaling.

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References

Shin S, Kogerman P, Lindstrom E, Toftgard R, Biesecker L . GLI3 mutations in human disorders mimic Drosophila cubitus interruptus protein functions and localization. Proc Natl Acad Sci U S A. 1999; 96(6):2880-4. PMC: 15863. DOI: 10.1073/pnas.96.6.2880. View

Rape M, Jentsch S . Taking a bite: proteasomal protein processing. Nat Cell Biol. 2002; 4(5):E113-6. DOI: 10.1038/ncb0502-e113. View

Zhang W, Zhao Y, Tong C, Wang G, Wang B, Jia J . Hedgehog-regulated Costal2-kinase complexes control phosphorylation and proteolytic processing of Cubitus interruptus. Dev Cell. 2005; 8(2):267-78. DOI: 10.1016/j.devcel.2005.01.001. View

Eide E, Woolf M, Kang H, Woolf P, Hurst W, Camacho F . Control of mammalian circadian rhythm by CKIepsilon-regulated proteasome-mediated PER2 degradation. Mol Cell Biol. 2005; 25(7):2795-807. PMC: 1061645. DOI: 10.1128/MCB.25.7.2795-2807.2005. View

Kanemori Y, Uto K, Sagata N . Beta-TrCP recognizes a previously undescribed nonphosphorylated destruction motif in Cdc25A and Cdc25B phosphatases. Proc Natl Acad Sci U S A. 2005; 102(18):6279-84. PMC: 1083676. DOI: 10.1073/pnas.0501873102. View

Jia J, Zhang L, Zhang Q, Tong C, Wang B, Hou F . Phosphorylation by double-time/CKIepsilon and CKIalpha targets cubitus interruptus for Slimb/beta-TRCP-mediated proteolytic processing. Dev Cell. 2005; 9(6):819-30. DOI: 10.1016/j.devcel.2005.10.006. View

Stone D, Murone M, Luoh S, Ye W, Armanini M, Gurney A . Characterization of the human suppressor of fused, a negative regulator of the zinc-finger transcription factor Gli. J Cell Sci. 1999; 112 ( Pt 23):4437-48. DOI: 10.1242/jcs.112.23.4437. View

Wang B, Fallon J, Beachy P . Hedgehog-regulated processing of Gli3 produces an anterior/posterior repressor gradient in the developing vertebrate limb. Cell. 2000; 100(4):423-34. DOI: 10.1016/s0092-8674(00)80678-9. View

Methot N, Basler K . Suppressor of fused opposes hedgehog signal transduction by impeding nuclear accumulation of the activator form of Cubitus interruptus. Development. 2000; 127(18):4001-10. DOI: 10.1242/dev.127.18.4001. View

10.

Lin L, DeMartino G, Greene W . Cotranslational dimerization of the Rel homology domain of NF-kappaB1 generates p50-p105 heterodimers and is required for effective p50 production. EMBO J. 2000; 19(17):4712-22. PMC: 302078. DOI: 10.1093/emboj/19.17.4712. View

11.

Ciechanover A, Gonen H, Bercovich B, Cohen S, Fajerman I, Israel A . Mechanisms of ubiquitin-mediated, limited processing of the NF-kappaB1 precursor protein p105. Biochimie. 2001; 83(3-4):341-9. DOI: 10.1016/s0300-9084(01)01239-1. View

12.

Jia J, Amanai K, Wang G, Tang J, Wang B, Jiang J . Shaggy/GSK3 antagonizes Hedgehog signalling by regulating Cubitus interruptus. Nature. 2002; 416(6880):548-52. DOI: 10.1038/nature733. View

13.

Amit S, Hatzubai A, Birman Y, Andersen J, Ben-Shushan E, Mann M . Axin-mediated CKI phosphorylation of beta-catenin at Ser 45: a molecular switch for the Wnt pathway. Genes Dev. 2002; 16(9):1066-76. PMC: 186245. DOI: 10.1101/gad.230302. View

14.

Noureddine M, Donaldson T, Thacker S, Duronio R . Drosophila Roc1a encodes a RING-H2 protein with a unique function in processing the Hh signal transducer Ci by the SCF E3 ubiquitin ligase. Dev Cell. 2002; 2(6):757-70. DOI: 10.1016/s1534-5807(02)00164-8. View

15.

Fong A, Sun S . Genetic evidence for the essential role of beta-transducin repeat-containing protein in the inducible processing of NF-kappa B2/p100. J Biol Chem. 2002; 277(25):22111-4. DOI: 10.1074/jbc.C200151200. View

16.

Taylor M, Liu L, Raffel C, Hui C, Mainprize T, Zhang X . Mutations in SUFU predispose to medulloblastoma. Nat Genet. 2002; 31(3):306-10. DOI: 10.1038/ng916. View

17.

Ou C, Lin Y, Chen Y, Chien C . Distinct protein degradation mechanisms mediated by Cul1 and Cul3 controlling Ci stability in Drosophila eye development. Genes Dev. 2002; 16(18):2403-14. PMC: 187440. DOI: 10.1101/gad.1011402. View

18.

McMahon A, Ingham P, Tabin C . Developmental roles and clinical significance of hedgehog signaling. Curr Top Dev Biol. 2003; 53:1-114. DOI: 10.1016/s0070-2153(03)53002-2. View

19.

Liu C, Corboy M, DeMartino G, Thomas P . Endoproteolytic activity of the proteasome. Science. 2002; 299(5605):408-11. PMC: 3516294. DOI: 10.1126/science.1079293. View

20.

Orlicky S, Tang X, Willems A, Tyers M, Sicheri F . Structural basis for phosphodependent substrate selection and orientation by the SCFCdc4 ubiquitin ligase. Cell. 2003; 112(2):243-56. DOI: 10.1016/s0092-8674(03)00034-5. View