title: Generation and quantitative proteomics analysis of CK2α/α'(-/-)cells.
abstract: CK2 is a ubiquitous, constitutively active, highly pleiotropic, acidophilic Ser/Thr protein kinase whose holoenzyme is composed of two catalytic (α and/or α') subunits and a dimer of a non-catalytic β subunit. Abnormally high CK2 level/activity is often associated with malignancy and a variety of cancer cells have been shown to rely on it to escape apoptosis. To gain information about the actual "druggability" of CK2 and to dissect CK2 dependent cellular processes that are instrumental to the establishment and progression of neoplasia we have exploited the CRISPR/Cas9 genome editing technology to generate viable clones of C2C12 myoblasts devoid of either both the CK2 catalytic subunits or its regulatory β-subunit. Suppression of both CK2 catalytic subunits promotes the disappearance of the β-subunit as well, through its accelerated proteasomal degradation. A quantitative proteomics analysis of CK2α/α'(-/-)versus wild type cells shows that knocking out both CK2 catalytic subunits causes a rearrangement of the proteomics profile, with substantially altered level ( > 50%) of 240 proteins, 126 of which are up-regulated, while the other are down-regulated. A functional analysis reveals that up- and down-regulated proteins tend to be segregated into distinct sub-cellular compartments and play different biological roles, consistent with a global rewiring underwent by the cell to cope with the lack of CK2.
authors.affiliation: Department of Biomedical Sciences, University of Padova, Via U. Bassi 58/B, Padova, Italy.
authors.forename: Christian
authors.initials: C
authors.lastname: Borgo
authors.affiliation: Department of Biomedical Sciences, University of Padova, Via U. Bassi 58/B, Padova, Italy.|Proteomics Center, University of Padova and Azienda Ospedaliera di Padova, via G. Orus 2/B, Padova, Italy.
authors.forename: Cinzia
authors.initials: C
authors.lastname: Franchin
authors.affiliation: Proteomics Center, University of Padova and Azienda Ospedaliera di Padova, via G. Orus 2/B, Padova, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Scalco
authors.affiliation: Department of Molecular Medicine, University of Padova, Via Gabelli 63, Padova, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Bosello-Travain
authors.affiliation: Department of Biomedical Sciences, University of Padova, Via U. Bassi 58/B, Padova, Italy.
authors.forename: Arianna
authors.initials: A
authors.lastname: Donella-Deana
authors.affiliation: Department of Biomedical Sciences, University of Padova, Via U. Bassi 58/B, Padova, Italy.|Proteomics Center, University of Padova and Azienda Ospedaliera di Padova, via G. Orus 2/B, Padova, Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Arrigoni
authors.affiliation: Department of Biomedical Sciences, University of Padova, Via U. Bassi 58/B, Padova, Italy.
authors.forename: Mauro
authors.initials: M
authors.lastname: Salvi
authors.affiliation: Department of Biomedical Sciences, University of Padova, Via U. Bassi 58/B, Padova, Italy.|CNR Institute of Neurosciences, Via U. Bassi 58/B, Padova, Italy.
authors.forename: Lorenzo A
authors.initials: LA
authors.lastname: Pinna
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D047390:Casein Kinase II / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002384:Catalysis; D020134:Catalytic Domain; D002460:Cell Line; D002853:Chromatography, Liquid; D000072669:Gene Editing; D015870:Gene Expression; D055786:Gene Knockout Techniques; D051379:Mice; D021122:Protein Subunits; D021381:Protein Transport; D020543:Proteome; D040901:Proteomics / Q000379:methods; D053719:Tandem Mass Spectrometry
chemical_list: D021122:Protein Subunits; D020543:Proteome; D047390:Casein Kinase II