Dataset Information

CRISPR/Cas9-mediated knockout of the ubiquitin variant UbKEKS reveals a role in regulating nucleolar structures and composition

ABSTRACT: Ubiquitination is a well-known post-translational modification responsible for one of the most powerful multi-layered communication and regulation system in the cell. Over the past decades, new ubiquitin variants and ubiquitin-like proteins arose to further enrich this mechanism. Among them, the recently discovered ubiquitin variant UbKEKS can specifically target many proteins and yet, functional consequences of this new modification remain unknown. The absence of UbKEKS induces accumulation of lamin A in the nucleoli, highlighting the need for deeper investigations about protein composition and functions regulation of this highly dynamic and membrane-less compartment. By using data independent acquisition mass spectrometry and microscopy, we show here that despite not impacting protein stability, UbKEKS is required for correct nucleolar organization. The absence of UbKEKS increases nucleoli’s size and accentuate their circularity while disrupting dense fibrillar component and fibrillar center structures. Moreover, depletion of UbKEKS leads to distinct changes in nucleolar composition. Notably, lack of UbKEKS favors nucleolar sequestration of known apoptotic regulators such as IFI16 or P14arf. Increase of apoptosis in UbKEKS knockout cells was later validated by flow cytometry and real-time cellular growth monitoring. In the end, this work identifies the first cellular functions of the UbKEKS variant and lay the foundation stone to establish UbKEKS as a new universal layer of regulation in the already complex ubiquitination system.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture, Early Embryonic Cell, Fibroblast, Hela Cell

DISEASE(S): Colon Cancer

SUBMITTER: Dominique Levesque

LAB HEAD: Francois-Michel Boisvert

PROVIDER: PXD040778 | Pride | 2023-10-11

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	DIA_Julie_Hela_27_N1a_9974.d.7z	Other
	DIA_Julie_Hela_27_N1b_10643.d.7z	Other
	DIA_Julie_Hela_27_N1c_10644.d.7z	Other
	DIA_Julie_Hela_27_N2a_10645.d.7z	Other
	DIA_Julie_Hela_27_N2b_10646.d.7z	Other

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CRISPR/Cas9-mediated knockout of the ubiquitin variant UbKEKS reveals a role in regulating nucleolar structures and composition

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