Hedgehog-regulated ubiquitination controls smoothened trafficking and cell surface expression in Drosophila

Shuang Li, Yongbin Chen, Qing Shi, Tao Yue, Bing Wang, Jin Jiang

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

Hedgehog transduces signal by promoting cell surface expression of the seven-transmembrane protein Smoothened (Smo) in Drosophila, but the underlying mechanism remains unknown. Here we demonstrate that Smo is downregulated by ubiquitin-mediated endocytosis and degradation, and that Hh increases Smo cell surface expression by inhibiting its ubiquitination. We find that Smo is ubiquitinated at multiple Lysine residues including those in its autoinhibitory domain (SAID), leading to endocytosis and degradation of Smo by both lysosome- and proteasome-dependent mechanisms. Hh inhibits Smo ubiquitination via PKA/CK1-mediated phosphorylation of SAID, leading to Smo cell surface accumulation. Inactivation of the ubiquitin activating enzyme Uba1 or perturbation of multiple components of the endocytic machinery leads to Smo accumulation and Hh pathway activation. In addition, we find that the non-visual β-arrestin Kurtz (Krz) interacts with Smo and acts in parallel with ubiquitination to downregulate Smo. Finally, we show that Smo ubiquitination is counteracted by the deubiquitinating enzyme UBPY/USP8. Gain and loss of UBPY lead to reciprocal changes in Smo cell surface expression. Taken together, our results suggest that ubiquitination plays a key role in the downregulation of Smo to keep Hh pathway activity off in the absence of the ligand, and that Hh-induced phosphorylation promotes Smo cell surface accumulation by inhibiting its ubiquitination, which contributes to Hh pathway activation.

Original languageEnglish (US)
Article numbere1001239
JournalPLoS Biology
Volume10
Issue number1
DOIs
StatePublished - Jan 2012

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Hedgehogs
Erinaceidae
Ubiquitination
Drosophila
Phosphorylation
Down-Regulation
endocytosis
cells
Endocytosis
Ubiquitin-Activating Enzymes
Chemical activation
phosphorylation
Arrestin
Degradation
arrestins
Proteasome Endopeptidase Complex
Ubiquitin
ubiquitin-protein ligase
transmembrane proteins
degradation

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

Hedgehog-regulated ubiquitination controls smoothened trafficking and cell surface expression in Drosophila. / Li, Shuang; Chen, Yongbin; Shi, Qing; Yue, Tao; Wang, Bing; Jiang, Jin.

In: PLoS Biology, Vol. 10, No. 1, e1001239, 01.2012.

Research output: Contribution to journalArticle

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