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/ Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. imex curation Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions 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 Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es Accepted 2009-FEB-14 by DAVET. 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions imex curation Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. imex curation Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions 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 Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es Accepted 2009-FEB-14 by DAVET. 03-Mar-2009: Contacted by JYOTI. davet Only protein-protein interactions imex curation Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. imex curation Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions 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 tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es Accepted 2009-FEB-14 by DAVET. 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions imex curation Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. imex curation Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions 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. confocal microscopy confocal microscopy monoclonal immunost monoclonal antibody immunostaining monoclonal immunost Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es Accepted 2009-FEB-14 by DAVET. 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions imex curation Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. imex curation Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions 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 anti tag western anti tag western blot anti tag western Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es Accepted 2009-FEB-14 by DAVET. 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions imex curation Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. imex curation Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es Accepted 2009-FEB-14 by DAVET. 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions imex curation Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. imex curation Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions in vitro In vitro pull down pull down fingerprinting peptide massfingerprinting fingerprinting Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es Accepted 2009-FEB-14 by DAVET. 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions imex curation Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. imex curation Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 confocal microscopy confocal microscopy immunostaining immunostaining Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es Accepted 2009-FEB-14 by DAVET. 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions imex curation Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. imex curation Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay autoradiography autoradiography Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es Accepted 2009-FEB-14 by DAVET. 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions imex curation Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. Plk3 interacts and specifically phosphorylates VRK1 in Ser342, a downstream target in a pathway that induces Golgi fragmentation. imex curation Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions human-a_549 Homo sapiens Caucasian lung carcinoma cell line a_549 Human Caucasian lung carcinoma confocal microscopy confocal microscopy monoclonal immunost monoclonal antibody immunostaining monoclonal immunost Lopez-Sanchez I, Sanz-Garcia M, and Lazo PA Apoptosis - Interactions involving proteins with a function related to apoptosis 2008-07-22: Lazo P, Instituto de Biologia Molecular y Celular del Cancer, Centro de Investigacion del Cancer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain plazozbi@usal.es Accepted 2009-FEB-14 by DAVET. 03-Feb-2009: Contacted by JYOTI. davet Only protein-protein interactions imex curation p53_mouse Cellular tumor antigen p53 Tp53 P53 Trp53 Tumor suppressor p53 protein protein mouse Mus musculus Mouse MTAMEESQSDISLELPLSQETFSGLWKLLPPEDILPSPHCMDDLLLPQDVEEFFEGPSEALRVSGAPAAQDPVTETPGPVAPAPATPWPLSSFVPSQKTYQGNYGFHLGFLQSGTAKSVMCTYSPPLNKLFCQLAKTCPVQLWVSATPPAGSRVRAMAIYKKSQHMTEVVRRCPHHERCSDGDGLAPPQHLIRVEGNLYPEYLEDRQTFRHSVVVPYEPPEAGSEYTTIHYKYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRDSFEVRVCACPGRDRRTEEENFRKKEVLCPELPPGSAKRALPTCTSASPPQKKKPLDGEYFTLKIRGRKRFEMFRELNEALELKDAHATEESGDSRAHSSYLKTKKGQSTSRHKKTMVKKVGPDSD DIP protein P02340 original sequence version: 117 DIP protein P02340 original sequence version: 165 DIP protein P02340 original sequence version: 187 DIP protein P02340 original sequence version: 201 DIP protein P02340 original sequence version: 111 DIP protein P02340 original sequence version: 141 DIP protein P02340 original sequence version: 180 DIP protein P02340 original sequence version: 163 DIP protein P02340 original sequence version: 152 ED5A607378D921D7 plk3_human Serine/threonine-protein kinase PLK3 Polo-like kinase 3 Cytokine-inducible serine/threonine-protein kinase FGF-inducible kinase Proliferation-related kinase PLK3 CNK FNK PRK protein protein human Homo sapiens Human MEPAAGFLSPRPFQRAAAAPAPPAGPGPPPSALRGPELEMLAGLPTSDPGRLITDPRSGRTYLKGRLLGKGGFARCYEATDTETGSAYAVKVIPQSRVAKPHQREKILNEIELHRDLQHRHIVRFSHHFEDADNIYIFLELCSRKSLAHIWKARHTLLEPEVRYYLRQILSGLKYLHQRGILHRDLKLGNFFITENMELKVGDFGLAARLEPPEQRKKTICGTPNYVAPEVLLRQGHGPEADVWSLGCVMYTLLCGSPPFETADLKETYRCIKQVHYTLPASLSLPARQLLAAILRASPRDRPSIDQILRHDFFTKGYTPDRLPISSCVTVPDLTPPNPARSLFAKVTKSLFGRKKKSKNHAQERDEVSGLVSGLMRTSVGHQDARPEAPAASGPAPVSLVETAPEDSSPRGTLASSGDGFEEGLTVATVVESALCALRNCIAFMPPAEQNPAPLAQPEPLVWVSKWVDYSNKFGFGYQLSSRRVAVLFNDGTHMALSANRKTVHYNPTSTKHFSFSVGAVPRALQPQLGILRYFASYMEQHLMKGGDLPSVEEVEVPAPPLLLQWVKTDQALLMLFSDGTVQVNFYGDHTKLILSGWEPLLVTFVARNRSACTYLASHLRQLGCSPDLRQRLRYALRLLRDRSPA 324CA3E9DE482514 vrk1_human Serine/threonine-protein kinase VRK1 Vaccinia-related kinase 1 VRK1 protein protein human Homo sapiens Human MPRVKAAQAGRQSSAKRHLAEQFAVGEIITDMAKKEWKVGLPIGQGGFGCIYLADMNSSESVGSDAPCVVKVEPSDNGPLFTELKFYQRAAKPEQIQKWIRTRKLKYLGVPKYWGSGLHDKNGKSYRFMIMDRFGSDLQKIYEANAKRFSRKTVLQLSLRILDILEYIHEHEYVHGDIKASNLLLNYKNPDQVYLVDYGLAYRYCPEGVHKEYKEDPKRCHDGTIEFTSIDAHNGVAPSRRGDLEILGYCMIQWLTGHLPWEDNLKDPKYVRDSKIRYRENIASLMDKCFPEKNKPGEIAKYMETVKLLDYTEKPLYENLRDILLQGLKAIGSKDDGKLDLSVVENGGLKAKTITKKRKKEIEESKEPGVEDTEWSNTQTEEAIQTRSRTRKRVQK 5640C624BF059949 gogb1_human Golgin subfamily B member 1 Giantin Macrogolgin 372 kDa Golgi complex-associated protein GOLGB1 protein protein human Homo sapiens Human MLSRLSGLANVVLHELSGDDDTDQNMRAPLDPELHQESDMEFNNTTQEDVQERLAYAEQLVVELKDIIRQKDVQLQQKDEALQEERKAADNKIKKLKLHAKAKLTSLNKYIEEMKAQGGTVLPTEPQSEEQLSKHDKSSTEEEMEIEKIKHKLQEKEELISTLQAQLTQAQAEQPAQSSTEMEEFVMMKQQLQEKEEFISTLQAQLSQTQAEQAAQQVVREKDARFETQVRLHEDELLQLVTQADVETEMQQKLRVLQRKLEEHEESLVGRAQVVDLLQQELTAAEQRNQILSQQLQQMEAEHNTLRNTVETEREESKILLEKMELEVAERKLSFHNLQEEMHHLLEQFEQAGQAQAELESRYSALEQKHKAEMEEKTSHILSLQKTGQELQSACDALKDQNSKLLQDKNEQAVQSAQTIQQLEDQLQQKSKEISQFLNRLPLQQHETASQTSFPDVYNEGTQAVTEENIASLQKRVVELENEKGALLLSSIELEELKAENEKLSSQITLLEAQNRTGEADREVSEISIVDIANKRSSSAEESGQDVLENTFSQKHKELSVLLLEMKEAQEEIAFLKLQLQGKRAEEADHEVLDQKEMKQMEGEGIAPIKMKVFLEDTGQDFPLMPNEESSLPAVEKEQASTEHQSRTSEEISLNDAGVELKSTKQDGDKSLSAVPDIGQCHQDELERLKSQILELELNFHKAQEIYEKNLDEKAKEISNLNQLIEEFKKNADNNSSAFTALSEERDQLLSQVKELSMVTELRAQVKQLEMNLAEAERQRRLDYESQTAHDNLLTEQIHSLSIEAKSKDVKIEVLQNELDDVQLQFSEQSTLIRSLQSQLQNKESEVLEGAERVRHISSKVEELSQALSQKELEITKMDQLLLEKKRDVETLQQTIEEKDQQVTEISFSMTEKMVQLNEEKFSLGVEIKTLKEQLNLLSRAEEAKKEQVEEDNEVSSGLKQNYDEMSPAGQISKEELQHEFDLLKKENEQRKRKLQAALINRKELLQRVSRLEEELANLKDESKKEIPLSETERGEVEEDKENKEYSEKCVTSKCQEIEIYLKQTISEKEVELQHIRKDLEEKLAAEEQFQALVKQMNQTLQDKTNQIDLLQAEISENQAIIQKLITSNTDASDGDSVALVKETVVISPPCTGSSEHWKPELEEKILALEKEKEQLQKKLQEALTSRKAILKKAQEKERHLREELKQQKDDYNRLQEQFDEQSKENENIGDQLRQLQIQVRESIDGKLPSTDQQESCSSTPGLEEPLFKATEQHHTQPVLESNLCPDWPSHSEDASALQGGTSVAQIKAQLKEIEAEKVELELKVSSTTSELTKKSEEVFQLQEQINKQGLEIESLKTVSHEAEVHAESLQQKLESSQLQIAGLEHLRELQPKLDELQKLISKKEEDVSYLSGQLSEKEAALTKIQTEIIEQEDLIKALHTQLEMQAKEHDERIKQLQVELCEMKQKPEEIGEESRAKQQIQRKLQAALISRKEALKENKSLQEELSLARGTIERLTKSLADVESQVSAQNKEKDTVLGRLALLQEERDKLITEMDRSLLENQSLSSSCESLKLALEGLTEDKEKLVKEIESLKSSKIAESTEWQEKHKELQKEYEILLQSYENVSNEAERIQHVVEAVRQEKQELYGKLRSTEANKKETEKQLQEAEQEMEEMKEKMRKFAKSKQQKILELEEENDRLRAEVHPAGDTAKECMETLLSSNASMKEELERVKMEYETLSKKFQSLMSEKDSLSEEVQDLKHQIEGNVSKQANLEATEKHDNQTNVTEEGTQSIPGETEEQDSLSMSTRPTCSESVPSAKSANPAVSKDFSSHDEINNYLQQIDQLKERIAGLEEEKQKNKEFSQTLENEKNTLLSQISTKDGELKMLQEEVTKMNLLNQQIQEELSRVTKLKETAEEEKDDLEERLMNQLAELNGSIGNYCQDVTDAQIKNELLESEMKNLKKCVSELEEEKQQLVKEKTKVESEIRKEYLEKIQGAQKEPGNKSHAKELQELLKEKQQEVKQLQKDCIRYQEKISALERTVKALEFVQTESQKDLEITKENLAQAVEHRKKAQAELASFKVLLDDTQSEAARVLADNLKLKKELQSNKESVKSQMKQKDEDLERRLEQAEEKHLKEKKNMQEKLDALRREKVHLEETIGEIQVTLNKKDKEVQQLQENLDSTVTQLAAFTKSMSSLQDDRDRVIDEAKKWERKFSDAIQSKEEEIRLKEDNCSVLKDQLRQMSIHMEELKINISRLEHDKQIWESKAQTEVQLQQKVCDTLQGENKELLSQLEETRHLYHSSQNELAKLESELKSLKDQLTDLSNSLEKCKEQKGNLEGIIRQQEADIQNSKFSYEQLETDLQASRELTSRLHEEINMKEQKIISLLSGKEEAIQVAIAELRQQHDKEIKELENLLSQEEEENIVLEEENKKAVDKTNQLMETLKTIKKENIQQKAQLDSFVKSMSSLQNDRDRIVGDYQQLEERHLSIILEKDQLIQEAAAENNKLKEEIRGLRSHMDDLNSENAKLDAELIQYREDLNQVITIKDSQQKQLLEVQLQQNKELENKYAKLEEKLKESEEANEDLRRSFNALQEEKQDLSKEIESLKVSISQLTRQVTALQEEGTLGLYHAQLKVKEEEVHRLSALFSSSQKRIAELEEELVCVQKEAAKKVGEIEDKLKKELKHLHHDAGIMRNETETAEERVAELARDLVEMEQKLLMVTKENKGLTAQIQSFGRSMSSLQNSRDHANEELDELKRKYDASLKELAQLKEQGLLNRERDALLSETAFSMNSTEENSLSHLEKLNQQLLSKDEQLLHLSSQLEDSYNQVQSFSKAMASLQNERDHLWNELEKFRKSEEGKQRSAAQPSTSPAEVQSLKKAMSSLQNDRDRLLKELKNLQQQYLQINQEITELHPLKAQLQEYQDKTKAFQIMQEELRQENLSWQHELHQLRMEKSSWEIHERRMKEQYLMAISDKDQQLSHLQNLIRELRSSSSQTQPLKVQYQRQASPETSASPDGSQNLVYETELLRTQLNDSLKEIHQKELRIQQLNSNFSQLLEEKNTLSIQLCDTSQSLRENQQHYGDLLNHCAVLEKQVQELQAGPLNIDVAPGAPQEKNGVHRKSDPEELREPQQSFSEAQQQLCNTRQEVNELRKLLEEERDQRVAAENALSVAEEQIRRLEHSEWDSSRTPIIGSCGTQEQALLIDLTSNSCRRTRSGVGWKRVLRSLCHSRTRVPLLAAIYFLMIHVLLILCFTGHL ED5D043C3A5C6432 atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) vrk1-plk3-1 7 12 unspecified role unspecified role prey prey region identified peptide identified peptide certain certain sequence position certain certain certain sequence position certain region identified peptide identified peptide certain certain sequence position certain certain certain sequence position certain region identified peptide identified peptide certain certain sequence position certain certain certain sequence position certain region identified peptide identified peptide certain certain sequence position certain certain certain sequence position certain 13 unspecified role unspecified role bait bait physical association physical association S2 This interaction between VRK1 and Plk3 was also detected by proteomic analysis of proteins associated with VRK1 in HEK293T cells (29). The proteomic identification by mass-spectrometry of Plk3 is shown in Fig. S2. vrk1-plk3-2 3 12 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 3a lysates were used for immunoprecipitation with either an anti-HA (Fig. 3A, bottom left) or anti-Flag antibody (Fig. 3A, bottom right), and the immunoprecipitates were used for immunoblotting with the reciprocal antibody. In both cases the VRK1-Plk3 interaction was detected transfection vrk1-plk3-3 4 12 unspecified role unspecified role neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 unspecified role unspecified role neutral component neutral component colocalization colocalization In the HEK293T cell line VRK1 has a particulate perinuclear, nuclear and fine granular cytosolic aspect and colocalizes with Plk3 in the perinuclear granules. ( 1 Fig.2C). 2C vrk1-plk3-4 1 12 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait physical association physical association 2B In the VRK1 immunoprecipitate the Plk3 protein was detectable (Fig. 2B). vrk1-plk3-5 9 15 enzyme target enzyme target substrate substrate unspecified role unspecified role 13 enzyme target enzyme target substrate substrate unspecified role unspecified role Q99986:p.Lys179Glu mutation mutation certain certain sequence position certain certain certain sequence position certain K E Wed May 02 19:06:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys179glu' Tue Jun 30 18:24:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys179Glu' region 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 ecoli Escherichia coli (strain K12) 12 enzyme enzyme unspecified role unspecified role region 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 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. phosphorylation phosphorylation reaction phosphorylation 1d Inactive VRK1K179E was phosphorylated when Plk3 was present (Fig.1D, lane 2), but not in the control Kinase dead VRK1 was used to show this interaction. PLK3 and VRK1 are both phosphorylated vrk1-plk3-6 9 12 enzyme enzyme unspecified role unspecified role region 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 ecoli Escherichia coli (strain K12) 13 enzyme target enzyme target substrate substrate unspecified role unspecified role Q99986:p.Lys179Glu mutation mutation certain certain sequence position certain certain certain sequence position certain K E Wed May 02 16:35:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys179glu' Tue Jun 30 17:18:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys179Glu' region 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 Q99986:p.Ser342Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain S A Wed May 02 18:37:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser342ala' Tue Jun 30 15:59:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser342Ala' ecoli Escherichia coli (strain K12) 15 enzyme target enzyme target substrate substrate unspecified role unspecified role phosphorylation phosphorylation reaction phosphorylation 1b When GSTVRK1 K179E/S342A 28 was used as substrate of Plk3 activity there was no phosphorylation signal of kinase-dead VRK1K179E/S342A (Fig. 1B, lane 5), suggesting that Ser342 is the target residue. Kinase dead VRK1 was used to show this interaction. PLK3 and VRK1 are both phosphorylated vrk1-2 9 15 enzyme target enzyme target substrate substrate unspecified role unspecified role 13 putative self putative self unspecified role unspecified role region 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 ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation 1c VRK1 is phosphorylated plk3-1 9 12 putative self putative self unspecified role unspecified role Q9H4B4:p.Lys91Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K R Wed May 02 18:50:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys91arg' Tue Jun 30 15:52:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys91Arg' region 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 ecoli Escherichia coli (strain K12) 15 enzyme target enzyme target substrate substrate unspecified role unspecified role phosphorylation phosphorylation reaction phosphorylation 1b PLK3 is phosphorylated Both kinases, Plk3 and VRK1, have autophosphorylation activity, as detected by the strong phosphorylation of the corresponding bands (Fig.1B, lanes 1 and 2). vrk1-1 9 15 enzyme target enzyme target substrate substrate unspecified role unspecified role 13 putative self putative self unspecified role unspecified role Q99986:p.Lys179Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K E Wed May 02 19:58:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys179glu' Tue Jun 30 17:31:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys179Glu' region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation 1b VRK1 is phosphorylated Both kinases, Plk3 and VRK1, have autophosphorylation activity, as detected by the strong phosphorylation of the corresponding bands (Fig.1B, lanes 1 and 2). vrk1-p53-3 9 13 enzyme enzyme unspecified role unspecified role region 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 ecoli Escherichia coli (strain K12) 11 enzyme target enzyme target substrate substrate unspecified role unspecified role region 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 4-88 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain The range was 1-85 but UniProt updated sequence so now range is 4-88. ecoli Escherichia coli (strain K12) 15 enzyme target enzyme target substrate substrate unspecified role unspecified role phosphorylation phosphorylation reaction phosphorylation 1c VRK1 and p53 are phosphorylated vrk1-plk3-7 10 13 unspecified role unspecified role neutral component neutral component 12 unspecified role unspecified role neutral component neutral component colocalization colocalization 2D Both VRK1 and Plk3 also colocalized in cytosolic granules in A549 cells (Fig. 2D) vrk1-giantin 8 14 unspecified role unspecified role neutral component neutral component 13 unspecified role unspecified role neutral component neutral component colocalization colocalization 5 In interphase and late telophase cells, VRK1 and giantin colocalized in a compacted body, but in intermediate steps along the progression of mitosis they retained their colocalization into smaller vesicles (Fig. 5). In interphase and late telophase cells, VRK1 and giantin colocalized in a compacted body, but in intermediate steps along the progression of mitosis they retained their colocalization into smaller golgi vesicles. vrk1-plk3-8 3 12 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 3a lysates were used for immunoprecipitation with either an anti-HA (Fig. 3A, bottom left) or anti-Flag antibody (Fig. 3A, bottom right), and the immunoprecipitates were used for immunoblotting with the reciprocal antibody. In both cases the VRK1-Plk3 interaction was detected transfection plk3-vrk1 5 13 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q99986:p.Lys179Glu mutation mutation certain certain sequence position certain certain certain sequence position certain K E Wed May 02 18:19:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys179glu' Tue Jun 30 15:38:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys179Glu' Q99986:p.Ser342Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain S A Wed May 02 15:03:41 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser342ala' Tue Jun 30 17:50:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser342Ala' 12 unspecified role unspecified role bait bait region 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 Q9H4B4:p.Lys91Arg mutation mutation certain certain sequence position certain certain certain sequence position certain K R Wed May 02 15:11:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys91arg' Tue Jun 30 19:13:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys91Arg' physical association physical association 3b VRK1 interacted with active and inactive Plk3; while the inactive VRK1K179E interacted more stably with inactive Plk3K52R . The interaction was lost if VRK1 has the S342A substitution, indicating that the intact target sequence in VRK1 was necessary in order to form a stable complex with Plk3 (Fig. 3B, top). VRK1 interacted with active and inactive PLK3. Inactive VRK1 lys179glu interacted with inactive PLK3 lys52arg. plk3-vrk1-1 2 13 unspecified role unspecified role prey prey 12 unspecified role unspecified role bait bait region 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 Q9H4B4:p.Lys91Arg mutation mutation certain certain sequence position certain certain certain sequence position certain K R Wed May 02 19:25:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys91arg' Tue Jun 30 16:42:38 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys91Arg' physical association physical association 3 Transfection of bait First the endogenous VRK1 was brought down from cells transfected with either pCEFL-GST-Plk3 or pCEFL-GST-Plk3K52R . In the pulldown VRK1 was detected associated to Plk3 but not to GST (Fig. 3C) plk3-vrk1-2 2 12 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait physical association physical association 3d Transfection of bait the endogenous Plk3 was detected bound to VRK1, active or kinase dead, but not to GST, in the extracts of cells transfected with pCEFLGSTVRK1 or pCEFLGSTVRK1K179E (Fig. 3D). plk3-vrk1-4 5 13 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region 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 12 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 4a The full-length, but not the C-terminal region of VRK1 was able to interact with PLK3. plk3-vrk1-5 5 12 unspecified role unspecified role bait bait region 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 13 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 4b,4c The common region of VRK1 that interacts with Plk3 is located in the region comprised between residues 1-267 (fig. 4C. vrk1-plk3-9 6 13 unspecified role unspecified role bait bait Q99986:p.Lys179Glu mutation mutation certain certain sequence position certain certain certain sequence position certain K E Wed May 02 16:06:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys179glu' Tue Jun 30 17:18:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys179Glu' region 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 ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role prey prey region 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction S1b