title: Insights into Gli Factors Ubiquitylation Methods.
abstract: The Hedgehog (Hh) signaling pathway governs cell growth and tissue development. Malfunctioning of several Hh pathway components, including the key transcriptional effector Gli proteins, is responsible for the onset of several tumors. Gli proteins activity is finely controlled by multilayered regulatory mechanisms, the most prominent of which is their proteasome-dependent proteolytic cleavage or massive ubiquitin-mediated proteolysis. Here, we described multiple procedures to determine whether a Gli protein is ubiquitylated both in a cellular context and in vitro, in basal conditions or by different E3 ubiquitin ligases and whether these processes are associated to Gli proteasome degradation.
authors.affiliation: Center for Life NanoScience at Sapienza, Istituto Italiano di Tecnologia, 00161, Rome, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Infante
authors.forename: Romina
authors.initials: R
authors.lastname: Alfonsi
authors.forename: Lucia
authors.initials: L
authors.lastname: Di Marcotullio
mesh_terms: D004789:Enzyme Activation; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D015513:Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins; D013379:Substrate Specificity; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism; D054875:Ubiquitination; D000071676:Zinc Finger Protein GLI1
chemical_list: D015513:Oncogene Proteins; D011994:Recombinant Proteins; D015534:Trans-Activators; D000071676:Zinc Finger Protein GLI1; D044767:Ubiquitin-Protein Ligases