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Ubiquiton – an inducible, linkage-specific polyubiquitylation tool

ABSTRACT: Here we introduce a set of engineered ubiquitin protein ligases and matching ubiquitin acceptor tags for the rapid, inducible linear (M1-), K48-, or K63-linked polyubiquitylation of proteins in yeast and mammalian cells. By applying the so-called ‘Ubiquiton’ (Ubo) system to proteasomal targeting and the endocytic pathway, we validate this tool for soluble cytoplasmic and nuclear as well as chromatin-associated and integral membrane proteins and demonstrate how it can be used to control the localization and stability of its targets. SILAC-based and label-free quantitative proteomics experiments were performed to verify the linkages of the polyubiquitin chains produced on a model substrate, GFP, in yeast and to identify potential off-target effects of a transgenic E3 system in human cells.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Homo Sapiens (human) Saccharomyces Cerevisiae (baker's Yeast)

TISSUE(S): Cell Culture

SUBMITTER: Jia-Xuan Chen

LAB HEAD: Helle Ulrich

PROVIDER: PXD045662 | Pride | 2023-12-15

REPOSITORIES: Pride

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			Action	DRS
	20230806_HU_0127_0129_E3_M1_MaxQuant_Output.zip	Other
	20230806_HU_0130_0132_E3_K48_MaxQuant_Output.zip	Other
	20230806_HU_0133_0135_E3_K63_MaxQuant_Output.zip	Other
	20230806_SFB1_IMB_HU_0127_hs_IP_SIL_150.raw	Raw
	20230806_SFB1_IMB_HU_0128_hs_IP_SIL_150.raw	Raw

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Ubiquiton – an inducible, linkage-specific polyubiquitylation tool

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