Reference Report

Reference
Citation	Xia, R., Jia, H., Fan, J., Liu, Y., Jia, J. (2012). USP8 promotes smoothened signaling by preventing its ubiquitination and changing its subcellular localization. PLoS Biol. 10(1): e1001238. (Export to RIS)
FlyBase ID	FBrf0218247
Publication Type	Research paper
PubMed ID	22253573
PubMed Abstract	The seven transmembrane protein Smoothened (Smo) is a critical component of the Hedgehog (Hh) signaling pathway and is regulated by phosphorylation, dimerization, and cell-surface accumulation upon Hh stimulation. However, it is not clear how Hh regulates Smo accumulation on the cell surface or how Hh regulates the intracellular trafficking of Smo. In addition, little is known about whether ubiquitination is involved in Smo regulation. In this study, we demonstrate that Smo is multi-monoubiquitinated and that Smo ubiquitination is inhibited by Hh and by phosphorylation. Using an in vivo RNAi screen, we identified ubiquitin-specific protease 8 (USP8) as a deubiquitinase that down-regulates Smo ubiquitination. Inactivation of USP8 increases Smo ubiquitination and attenuates Hh-induced Smo accumulation, leading to decreased Hh signaling activity. Moreover, overexpression of USP8 prevents Smo ubiquitination and elevates Smo accumulation, leading to increased Hh signaling activity. Mechanistically, we show that Hh promotes the interaction of USP8 with Smo aa625-753, which covers the three PKA and CK1 phosphorylation clusters. Finally, USP8 promotes the accumulation of Smo at the cell surface and prevents localization to the early endosomes, presumably by deubiquitinating Smo. Our studies identify USP8 as a positive regulator in Hh signaling by down-regulating Smo ubiquitination and thereby mediating Smo intracellular trafficking.
DOI	10.1371/journal.pbio.1001238
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Language of Publication	English
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Publication Type	Journal
Abbreviation	PLoS Biol.
Title	PLoS Biology
Publication Year	2003-
ISBN/ISSN	1545-7885 1544-9173
Data from Reference
Alleles (22)
	Pka-C1^E95 ptc⁹ ptc^H84 Scer\GAL4^Act5C.PU Scer\GAL4^ap-md544 Scer\GAL4^Bx-MS1096 shi^1.Scer\UAS shi^GD1529 smo^{Scer\UAS.attB.T:Hsap\MYC} smo^{Scer\UAS.J.T:Avic\GFP} smo^{Scer\UAS.J.T:Zzzz\FLAG} smo^{Scer\UAS.T:Hsap\MYC,T:Avic\GFP-CFP} smo^{Scer\UAS.T:Hsap\MYC,T:Uuuu\Ub} smo^{SD123.Scer\UAS.T:Avic\GFP-CFP} T:Avic\GFP^CFP Ubpy^{CS.Scer\UAS.T:Zzzz\FLAG} Ubpy^GD1285 Ubpy^KK100733 Ubpy^KO Ubpy^{NT3.CS.Scer\UAS.T:Ivir\HA1} Ubpy^{NT3.Scer\UAS.T:Ivir\HA1} Ubpy^{Scer\UAS.T:Zzzz\FLAG}
Constructs (14)
	P{Act5C-GAL4.U} P{GD1285} P{GD1529} P{KK100733} P{UAS-shi^ts1.K} P{UAS-smo.Fg} P{UAS-smo.GFP} PBac{UAS-Flag-USP8.CS} PBac{UAS-Flag-USP8} PBac{UAS-HA-USP8.NT3.CS} PBac{UAS-HA-USP8.NT3} PBac{UAS-Myc-smo.CFP} PBac{UAS-Myc-smo.Ub} PBac{UAS-smo.Myc}
Genes (26)
	CG4165 CG5384 CG7288 CG12082 CYLD dpp ec en faf hh not Pka-C1 ptc Rab5 Rab7 Scer\GAL4 scny shi smo T:Hsap\MYC T:Ivir\HA1 T:Uuuu\Ub T:Zzzz\FLAG trbd Ubpy Usp7
Insertions (2)
	P{GawB}ap^md544 P{GawB}Bx^MS1096
Natural transposons (1)
	Tni\piggyBac