@inbook{f0ce6456797240378cc0aa729bfd53f7,
title = "Cell-Based Assays for Smoothened Ubiquitination and Sumoylation",
abstract = "The Hedgehog (Hh) family of secreted proteins governs embryonic development and adult tissue homeostasis by regulating the abundance, localization, and activity of the GPCR family protein Smoothened (Smo). Smo trafficking and subcellular accumulation are controlled by multiple posttranslational modifications (PTMs) including phosphorylation, ubiquitination, and sumoylation, which appears to be conserved from Drosophila to mammals. Smo ubiquitination is dynamically regulated by E3 ubiquitin ligases and deubiquitinases (dubs) and is opposed by Hh signaling. By contrast, Smo sumoylation is stimulated by Hh, which counteracts Smo ubiquitination by recruiting the dub USP8. We describe cell-base assays for Smo ubiquitination and its regulation by Hh and the E3 ligases in Drosophila. We also describe assays for Smo sumoylation in both Drosophila and mammalian cultured cells.",
keywords = "Cul4, DDB1, Gβ, Hedgehog, Nedd4, Smo, Smurf, Sumoylation, USP8, Ubiquitination, Ulp1",
author = "Yuhong Han and Jin Jiang",
note = "Funding Information: This work was supported by grants from the National Institutes of Health (R35GM118063) and Welch Foundation (I-1603) to J.J. Publisher Copyright: {\textcopyright} 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2022",
doi = "10.1007/978-1-0716-1701-4_12",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "139--147",
booktitle = "Methods in Molecular Biology",
}