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/ boutet-2007-2 Regulation of Pax3 by proteasomal degradation of monoubiquitinated protein in skeletal muscle progenitors. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Boutet SC., Disatnik MH., Chan LS., Iori K., Rando TA. The reference Brooks et al. (2006) Molecular Cell, Volume 21(3), 307-315 (PMID 16455486) indicates that the ubiquitin construct was of human origin. rando@stanford.edu Cell (0092-8674) 2007 Only protein-protein interactions 11 imex curation boutet-2007-1 Regulation of Pax3 by proteasomal degradation of monoubiquitinated protein in skeletal muscle progenitors. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Boutet SC., Disatnik MH., Chan LS., Iori K., Rando TA. The reference Brooks et al. (2006) Molecular Cell, Volume 21(3), 307-315 (PMID 16455486) indicates that the ubiquitin construct was of human origin. rando@stanford.edu Cell (0092-8674) 2007 Only protein-protein interactions 11 imex curation boutet-2007-3 Regulation of Pax3 by proteasomal degradation of monoubiquitinated protein in skeletal muscle progenitors. mouse-c2c12 Mouse C3H muscle myoblast c2c12 Mouse C3H muscle myoblast anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Boutet SC., Disatnik MH., Chan LS., Iori K., Rando TA. The reference Brooks et al. (2006) Molecular Cell, Volume 21(3), 307-315 (PMID 16455486) indicates that the ubiquitin construct was of human origin. rando@stanford.edu Cell (0092-8674) 2007 Only protein-protein interactions 11 imex curation rd23b_mouse UV excision repair protein RAD23 homolog B XP-C repair-complementing complex 58 kDa protein Rad23b Mhr23b protein protein mouse Mus musculus Mouse MQVTLKTLQQQTFKIDIDPEETVKALKEKIESEKGKDAFPVAGQKLIYAGKILSDDTALKEYKIDEKNFVVVMVTKPKAVTTAVPATTQPSSTPSPTAVSSSPAVAAAQAPAPTPALPPTSTPASTAPASTTASSEPAPAGATQPEKPAEKPAQTPVLTSPAPADSTPGDSSRSNLFEDATSALVTGQSYENMVTEIMSMGYEREQVIAALRASFNNPDRAVEYLLMGIPGDRESQAVVDPPPQAVSTGTPQSPAVAAAAATTTATTTTTSGGHPLEFLRNQPQFQQMRQIIQQNPSLLPALLQQIGRENPQLLQQISQHQEHFIQMLNEPVQEAGSQGGGGGGGGGGGGGGGGGIAEAGSGHMNYIQVTPQEKEAIERLKALGFPEGLVIQAYFACEKNENLAANFLLQQNFDED rd23a_mouse UV excision repair protein RAD23 homolog A Rad23a Mhr23a protein protein mouse Mus musculus Mouse MAVTITLKTLQQQTFKIRMEPDETVKVLKEKIEAEKGRDAFPVAGQKLIYAGKILSDDVPIKEYHIDEKNFVVVMVTKAKAGQGIPAPPEASPTAVPEPSTPFPPVLASGMSHPPPTSREDKSPSEESTTTTSPESISGSVPSSGSSGREEDAASTLVTGSEYETMLTEIMSMGYERERVVAALRASYNNPHRAVEYLLTGIPGSPEPEHGSVQESQAPEQPATEAAGENPLEFLRDQPQFQNMRQVIQQNPALLPALLQQLGQENPQLLQQISRHQEQFIQMLNEPPGELADISDVEGEVGAIGEEAPQMNYIQVTPQEKEAIERLKALGFPESLVIQAYFACEKNENLAANFLLSQNFDDE pax3_mouse Paired box protein Pax-3 Pax3 Pax-3 protein protein mouse Mus musculus Mouse MTTLAGAVPRMMRPGPGQNYPRSGFPLEVSTPLGQGRVNQLGGVFINGRPLPNHIRHKIVEMAHHGIRPCVISRQLRVSHGCVSKILCRYQETGSIRPGAIGGSKPKQVTTPDVEKKIEEYKRENPGMFSWEIRDKLLKDAVCDRNTVPSVSSISRILRSKFGKGEEEEADLERKEAEESEKKAKHSIDGILSERASAPQSDEGSDIDSEPDLPLKRKQRRSRTTFTAEQLEELERAFERTHYPDIYTREELAQRAKLTEARVQVWFSNRRARWRKQAGANQLMAFNHLIPGGFPPTAMPTLPTYQLSETSYQPTSIPQAVSDPSSTVHRPQPLPPSTVHQSTIPSNADSSSAYCLPSTRHGFSSYTDSFVPPSGPSNPMNPTIGNGLSPQVMGLLTNHGGVPHQPQTDYALSPLTGGLEPTTTVSASCSQRLEHMKNVDSLPTSQPYCPPTYSTAGYSMDPVTGYQYGQYGQSKPWTF psmd4_human 26S proteasome non-ATPase regulatory subunit 4 PSMD4 MCB1 26S proteasome regulatory subunit S5A 26S proteasome regulatory subunit RPN10 Multiubiquitin chain-binding protein Antisecretory factor 1 protein protein human Homo sapiens Human MVLESTMVCVDNSEYMRNGDFLPTRLQAQQDAVNIVCHSKTRSNPENNVGLITLANDCEVLTTLTPDTGRILSKLHTVQPKGKITFCTGIRVAHLALKHRQGKNHKMRIIAFVGSPVEDNEKDLVKLAKRLKKEKVNVDIINFGEEEVNTEKLTAFVNTLNGKDGTGSHLVTVPPGPSLADALISSPILAGEGGAMLGLGASDFEFGVDPSADPELALALRVSMEEQRQRQEEEARRAAAASAAEAGIATTGTEDSDDALLKMTISQQEFGRTGLPDLSSMTEEEQIAYAMQMSLQGAEFGQAESADIDASSAMDTSEPAKEEDDYDVMQDPEFLQSVLENLPGVDPNNEAIRNAMGSLASQATKDGKKDKKEEDKK rd23b_human UV excision repair protein RAD23 homolog B XP-C repair-complementing complex 58 kDa protein RAD23B protein protein human Homo sapiens Human MQVTLKTLQQQTFKIDIDPEETVKALKEKIESEKGKDAFPVAGQKLIYAGKILNDDTALKEYKIDEKNFVVVMVTKPKAVSTPAPATTQQSAPASTTAVTSSTTTTVAQAPTPVPALAPTSTPASITPASATASSEPAPASAAKQEKPAEKPAETPVATSPTATDSTSGDSSRSNLFEDATSALVTGQSYENMVTEIMSMGYEREQVIAALRASFNNPDRAVEYLLMGIPGDRESQAVVDPPQAASTGAPQSSAVAAAAATTTATTTTTSSGGHPLEFLRNQPQFQQMRQIIQQNPSLLPALLQQIGRENPQLLQQISQHQEHFIQMLNEPVQEAGGQGGGGGGGSGGIAEAGSGHMNYIQVTPQEKEAIERLKALGFPEGLVIQAYFACEKNENLAANFLLQQNFDED ubb_human Polyubiquitin-B UBB protein protein human Homo sapiens Human MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGC cdn1a_mouse Cyclin-dependent kinase inhibitor 1 Cdkn1a Cip1 Waf1 p21 CDK-interacting protein 1 Melanoma differentiation-associated protein protein protein mouse Mus musculus Mouse MSNPGDVRPVPHRSKVCRCLFGPVDSEQLRRDCDALMAGCLQEARERWNFDFVTETPLEGNFVWERVRSLGLPKVYLSPGSRSRDDLGGDKRPSTSSALLQGPAPEDHVALSLSCTLVSERPEDSPGGPGTSQGRKRRQTSLTDFYHSKRRLVFCKRKP cdn1a_human Cyclin-dependent kinase inhibitor 1 CDKN1A CAP20 CDKN1 CIP1 MDA6 PIC1 SDI1 WAF1 p21 CDK-interacting protein 1 Melanoma differentiation-associated protein 6 protein protein human Homo sapiens Human MSEPAGDVRQNPCGSKACRRLFGPVDSEQLSRDCDALMAGCIQEARERWNFDFVTETPLEGDFAWERVRGLGLPKLYLPTGPRRGRDELGGGRRPGTSPALLQGTAEEDHVDLSLSCTLVPRSGEQAEGSPGGPGDSQGRKRRQTSMTDFYHSKRRLIFSKRKP psmd4-pax3 GST-S5a pulls down Pax3-GFP, Rad23A and Rad23B from mouse myoblast extracts 659219 N/A 659222 unspecified role unspecified role prey prey mouse-c2c12 Mouse C3H muscle myoblast c2c12 Mouse C3H muscle myoblast N/A 659223 unspecified role unspecified role prey prey mouse-c2c12 Mouse C3H muscle myoblast c2c12 Mouse C3H muscle myoblast N/A 659224 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false undefined mutation mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false undefined mutation mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false mouse-c2c12 Mouse C3H muscle myoblast c2c12 Mouse C3H muscle myoblast N/A 659225 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 false ecoli Escherichia coli (strain K12) association association false false false 5 A rad23b-pax3 hRAD23B UBA2 pulls down Pax3-GFP in vitro 659219 N/A 659226 unspecified role unspecified role bait bait c-terminal UBA2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false in vitro In vitro N/A 659224 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P24610:p.Lys475Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P24610:p.Lys437Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false mouse-c2c12 Mouse C3H muscle myoblast c2c12 Mouse C3H muscle myoblast physical association physical association false false false 5 B BAIT protein AAH20973 The mouse myoblasts were treated with MG132 to inhibit activity of the proteasome. psmd4-pax3-1 GST-S5a pulls down monoubiquitinated Pax3-GFP from extracts of mouse myoblasts 659219 N/A 659227 unspecified role unspecified role prey prey mouse-c2c12 Mouse C3H muscle myoblast c2c12 Mouse C3H muscle myoblast N/A 659224 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false mouse-c2c12 Mouse C3H muscle myoblast c2c12 Mouse C3H muscle myoblast N/A 659225 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 false ecoli Escherichia coli (strain K12) association association false false false 5 D The mouse myoblasts were treated with MG132 to block proteasome activity. PS5a-agarose pull-down was carried out followed by immuoprecipitation with a GFP antibody. Only a small fraction of immunoprecipitated Pax3 was monoubiquitinated, whereas virtually all Pax3 pulled down using S5a was monoubiquitinated. psmd4-pax3-2 GST-S5a pulls down Pax3-GFP from MEF extracts 659219 N/A 659225 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 false ecoli Escherichia coli (strain K12) N/A 659224 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false mouse-embryonic_fibroblast Mus musculus embryonic fibroblasts embryonic_fibroblast Embryonic fibroblast Fibroblasts taken from an embryonic as opposed to a foetal or adult source. Foetus is defined as the stage at which tissue diferentiation is initiated. Tissue taken from an embryonic as opposed to a foetal or adult source. Embryo is defined as the development stage prior to organ formation. physical association physical association false false false 6 A psmd4-pax3-3 GST-S5a pulls down polyubiquitinated p21 from extracts of mouse myoblasts 659219 N/A 659227 unspecified role unspecified role prey prey mouse-c2c12 Mouse C3H muscle myoblast c2c12 Mouse C3H muscle myoblast N/A 659228 unspecified role unspecified role prey prey mouse-c2c12 Mouse C3H muscle myoblast c2c12 Mouse C3H muscle myoblast N/A 659225 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 false ecoli Escherichia coli (strain K12) association association false false false The mouse myoblasts were treated with MG132 to block proteasome activity. S6 p21 coupled to four ubiquitin chains and conjugates with larger numbers of ubiquitin adducts was pulled down. rad23b-pax3-1 RAD23B pulls down Pax3-Ub in vitro 659219 N/A 659224 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) N/A 659226 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 false ecobb Escherichia coli (strain B / BL21) N/A 659227 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) association association false false false 5 C rad23b-pax3-2 RAD23B (UBA2) pulls down Pax3-Ub in vitro 659219 N/A 659224 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) N/A 659227 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) N/A 659226 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 false region UBA2 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) association association false false false 5 C rad23b-pax3-3 RAD23B (UBA2) pulls down Pax3 in vitro 659219 N/A 659226 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 false region UBA2 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 659224 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) physical association physical association false false false 5 C pax3-rps27a in an in vitro ubiquitination assay Pax3 is monoubiquitinated 659220 N/A 659224 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro Stoichiometry: 1.0 N/A 659227 unspecified role unspecified role prey prey Stoichiometry: 1.0 physical association physical association false false false 2 C, D, S4 Pax3 was also monoubiquitinated in the presence of ubiquitinKO (as exprected) and when His-tagged Ub or UbKO were used and Nickel-agarose purification carried out. P21 was polyubiquitinated when this assay was carried out, showing that all elements necessary for polyubiquitination were present. This interaction also occurred in the presence of a reaction solution containing 26S proteasome in the absence of MG132. cdkn1a-rps27a in an in vitro ubiquitination assay p21 is ubiquitinated 659220 N/A 659227 unspecified role unspecified role prey prey N/A 659229 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false 2 C pax3-rps27a-1 Pax3 is monoubiquitinated in myoblast extracts 659221 N/A 659224 unspecified role unspecified role bait bait region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 659227 unspecified role unspecified role prey prey Stoichiometry: 1.0 physical association physical association false false false 5 D P24610 transfection