Dataset Information

EGF-dependent ubiquitinome, phosphorylation and interactome analysis in control and CYLD-deficient cells

ABSTRACT: The goal of the present project was to investigate the EGF-dependent ubiquitination changes occurring in HeLa cells upon a CYLD deficiency. We applied SILAC-based quantitative mass spectrometry proteomics together with genetic engineering and biochemical approaches, in order to enrich the ubiquitinated proteins of the control and silenced cells in response to the growth factor. In order to get a deeper insight of the obtained results, we subsequently combined the enrichment of CYLD protein through immunoprecipitation with SILAC-based proteomics. This allowed us to decipher a specific tyrosine phosphorylation of CYLD that responds to the treatment with EGF and, therefore, we next aimed to analyse the ability of this modification to recruit potential interaction partners. We thus performed a peptide pull-down assay that uncovered a set of proteins specifically associated to the phosphorylated tyrosine and helped to uncover an unexpected, previously unreported role for this deubiquitinase.

INSTRUMENT(S): LTQ Orbitrap Velos, Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hela Cell

DISEASE(S): Cervix Carcinoma

SUBMITTER: VIRGINIA Sanchez-Quiles

LAB HEAD: BLAGOY BLAGOEV

PROVIDER: PXD003423 | Pride | 2017-06-05

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20130121_VSQ_qe2g120_011123r_IP_CYLD_1.raw	Raw
	20130121_VSQ_qe2g120_011123r_IP_CYLD_10.raw	Raw
	20130121_VSQ_qe2g120_011123r_IP_CYLD_11.raw	Raw
	20130121_VSQ_qe2g120_011123r_IP_CYLD_12.raw	Raw
	20130121_VSQ_qe2g120_011123r_IP_CYLD_2.raw	Raw

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Publications

Cylindromatosis Tumor Suppressor Protein (CYLD) Deubiquitinase is Necessary for Proper Ubiquitination and Degradation of the Epidermal Growth Factor Receptor.

Sanchez-Quiles Virginia V Akimov Vyacheslav V Osinalde Nerea N Francavilla Chiara C Puglia Michele M Barrio-Hernandez Inigo I Kratchmarova Irina I Olsen Jesper V JV Blagoev Blagoy B

Molecular & cellular proteomics : MCP 20170601 8

Cylindromatosis tumor suppressor protein (CYLD) is a deubiquitinase, best known as an essential negative regulator of the NFkB pathway. Previous studies have suggested an involvement of CYLD in epidermal growth factor (EGF)-dependent signal transduction as well, as it was found enriched within the tyrosine-phosphorylated complexes in cells stimulated with the growth factor. EGF receptor (EGFR) signaling participates in central cellular processes and its tight regulation, partly through ubiquitin ...[more]

PMID: 28572092

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