IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ http://www.ebi.ac.uk/ anonymous-2013a-1 Human VRK1 forms with p53 a preassembled stable basal complex that is activated in response to UV-induced DNA damage human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot 2013-01-30: P Lazo, Instituto de Biología Molecular y Celular del Cáncer, Centro de Investigación del Cáncer (CIC), Consejo Superior de Investigaciones Científicas (CSIC), Universidad de Salamanca. plazozbi@usal.es Only protein-protein interactions imex curation López-Sánchez I., Valbuena A., Vázquez-Cedeira M., Khadake J., Sanz-García M., Carrillo A., Lazo P.A. FEBS Lett. (0014-5793) 2014 anonymous-2013a-2 Human VRK1 forms with p53 a preassembled stable basal complex that is activated in response to UV-induced DNA damage human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. pull down pull down western blot western blot Immuno blot Immuno blot 2013-01-30: P Lazo, Instituto de Biología Molecular y Celular del Cáncer, Centro de Investigación del Cáncer (CIC), Consejo Superior de Investigaciones Científicas (CSIC), Universidad de Salamanca. plazozbi@usal.es Only protein-protein interactions imex curation López-Sánchez I., Valbuena A., Vázquez-Cedeira M., Khadake J., Sanz-García M., Carrillo A., Lazo P.A. FEBS Lett. (0014-5793) 2014 anonymous-2014k-3 Human VRK1 forms with p53 a preassembled stable basal complex that is activated in response to UV-induced DNA damage human-h1299 Homo sapiens lung lymph node carcinoma h1299 Human lung lymph node carcinoma p53 null. These cells have also been reported to secrete the peptide hormone neuromedin B (NMB), but not gastrin releasing peptide (GRP). Also known as NCI H1299 or CRL-5803. http://www.lgcpromochem.com/atcc/ ATCC number: CRL-5803. ATCC number of the cell line is CRL-5803. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot 2013-01-30: P Lazo, Instituto de Biología Molecular y Celular del Cáncer, Centro de Investigación del Cáncer (CIC), Consejo Superior de Investigaciones Científicas (CSIC), Universidad de Salamanca. plazozbi@usal.es Only protein-protein interactions imex curation López-Sánchez I., Valbuena A., Vázquez-Cedeira M., Khadake J., Sanz-García M., Carrillo A., Lazo P.A. FEBS Lett. (0014-5793) 2014 anonymous-2014k-1 Human VRK1 forms with p53 a preassembled stable basal complex that is activated in response to UV-induced DNA damage in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot 2013-01-30: P Lazo, Instituto de Biología Molecular y Celular del Cáncer, Centro de Investigación del Cáncer (CIC), Consejo Superior de Investigaciones Científicas (CSIC), Universidad de Salamanca. plazozbi@usal.es Only protein-protein interactions imex curation López-Sánchez I., Valbuena A., Vázquez-Cedeira M., Khadake J., Sanz-García M., Carrillo A., Lazo P.A. FEBS Lett. (0014-5793) 2014 anonymous-2014k-2 Human VRK1 forms with p53 a preassembled stable basal complex that is activated in response to UV-induced DNA damage human-h1299 Homo sapiens lung lymph node carcinoma h1299 Human lung lymph node carcinoma p53 null. These cells have also been reported to secrete the peptide hormone neuromedin B (NMB), but not gastrin releasing peptide (GRP). Also known as NCI H1299 or CRL-5803. http://www.lgcpromochem.com/atcc/ ATCC number: CRL-5803. ATCC number of the cell line is CRL-5803. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot 2013-01-30: P Lazo, Instituto de Biología Molecular y Celular del Cáncer, Centro de Investigación del Cáncer (CIC), Consejo Superior de Investigaciones Científicas (CSIC), Universidad de Salamanca. plazozbi@usal.es Only protein-protein interactions imex curation López-Sánchez I., Valbuena A., Vázquez-Cedeira M., Khadake J., Sanz-García M., Carrillo A., Lazo P.A. FEBS Lett. (0014-5793) 2014 vrk1_human Serine/threonine-protein kinase VRK1 Vaccinia-related kinase 1 VRK1 protein protein human Homo sapiens Human MPRVKAAQAGRQSSAKRHLAEQFAVGEIITDMAKKEWKVGLPIGQGGFGCIYLADMNSSESVGSDAPCVVKVEPSDNGPLFTELKFYQRAAKPEQIQKWIRTRKLKYLGVPKYWGSGLHDKNGKSYRFMIMDRFGSDLQKIYEANAKRFSRKTVLQLSLRILDILEYIHEHEYVHGDIKASNLLLNYKNPDQVYLVDYGLAYRYCPEGVHKEYKEDPKRCHDGTIEFTSIDAHNGVAPSRRGDLEILGYCMIQWLTGHLPWEDNLKDPKYVRDSKIRYRENIASLMDKCFPEKNKPGEIAKYMETVKLLDYTEKPLYENLRDILLQGLKAIGSKDDGKLDLSVVENGGLKAKTITKKRKKEIEESKEPGVEDTEWSNTQTEEAIQTRSRTRKRVQK p53_human Cellular tumor antigen p53 TP53 P53 Tumor suppressor p53 Phosphoprotein p53 Antigen NY-CO-13 protein protein human Homo sapiens Human MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLSPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPPEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMFRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD tp53-vrk1 1363310 n/a 1363315 unspecified role unspecified role prey prey n/a p53 1363316 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false 1A vrk1-tp53-4 1363310 n/a p53 1363316 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1363315 unspecified role unspecified role bait bait physical association physical association false false false 1A vrk1-tp53-3 1363311 n/a p53 1363316 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a VRK1 1363315 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q99986:p.Lys179Glu mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false f1b f1c f1d vrk1-tp53-5 1363311 n/a p53 1363316 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell P04637:p.Arg175His mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false n/a VRK1 1363315 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false f3a vrk1-tp53-7 1363312 n/a p53 1363316 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level P04637:p.Arg175His mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false n/a VRK1 1363315 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Q99986:p.Glu313Gly mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false Q99986:p.Glu313_Lys314delinsGlyIle mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false sf1 sf2 To further explore this interaction, several VRK1 phosphorylation mutants within the C-terminus (S342A, S342D, T355A, T355D, T378E, T390A and T390D) were also tested and found that none of them affected the VRK1-p53 interaction (Fig. S1). We also examined the effect of the double mutant VRK1 (E313G/K314I), a locally charged region of the VRK1 C-terminus, on the interaction (Fig. S2A and B) and we found that a change in the charge in this region resulted in increased binding. Noteworthy, when the mutation that causes spinal muscular atrophy, VRK1(R358X) [50], was tested we showed that VRK1 was still able to interact with p53 (Fig. S2A, B). vrk1-tp53-8 1363312 n/a VRK1 1363315 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Q99986:p.Glu313_Lys314delinsGlyIle mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false n/a p53 1363316 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false sf1 sf2 To further explore this interaction, several VRK1 phosphorylation mutants within the C-terminus (S342A, S342D, T355A, T355D, T378E, T390A and T390D) were also tested and found that none of them affected the VRK1-p53 interaction (Fig. S1). We also examined the effect of the double mutant VRK1 (E313G/K314I), a locally charged region of the VRK1 C-terminus, on the interaction (Fig. S2A and B) and we found that a change in the charge in this region resulted in increased binding. Noteworthy, when the mutation that causes spinal muscular atrophy, VRK1(R358X) [50], was tested we showed that VRK1 was still able to interact with p53 (Fig. S2A, B). tp53-vrk1-1 1363313 n/a p53 1363316 weight by staining molecular weight estimation by staining weight by staining unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a VRK1 1363315 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. physical association physical association false false false f2 vrk1-tp53-6 1363314 n/a p53 1363316 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell P04637:p.Arg175His mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false n/a VRK1 1363315 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false f3b