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/ kong-2010-2 RNF220, an E3 ubiquitin ligase that targets Sin3B for ubiquitination. 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 Kong Q., Zeng W., Wu J., Hu W., Li C., Mao B. Apoptosis - Interactions involving proteins with a function related to apoptosis Ubiquitination assay. mao@mail.kiz.ac.cn Biochem. Biophys. Res. Commun. (0006-291X) 2010 Only protein-protein interactions imex curation kong-2010-1 RNF220, an E3 ubiquitin ligase that targets Sin3B for ubiquitination. in vitro In vitro enzymatic study enzymatic study predetermined predetermined participant predetermined Kong Q., Zeng W., Wu J., Hu W., Li C., Mao B. Apoptosis - Interactions involving proteins with a function related to apoptosis Ubiquitination assay. mao@mail.kiz.ac.cn Biochem. Biophys. Res. Commun. (0006-291X) 2010 Only protein-protein interactions imex curation kong-2010-3 RNF220, an E3 ubiquitin ligase that targets Sin3B for ubiquitination. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Kong Q., Zeng W., Wu J., Hu W., Li C., Mao B. Apoptosis - Interactions involving proteins with a function related to apoptosis Ubiquitination assay. mao@mail.kiz.ac.cn Mouse 11 day embryo cDNA library (Clontech) Biochem. Biophys. Res. Commun. (0006-291X) 2010 Only protein-protein interactions imex curation kong-2010-4 RNF220, an E3 ubiquitin ligase that targets Sin3B for ubiquitination. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Kong Q., Zeng W., Wu J., Hu W., Li C., Mao B. Apoptosis - Interactions involving proteins with a function related to apoptosis Ubiquitination assay. mao@mail.kiz.ac.cn Biochem. Biophys. Res. Commun. (0006-291X) 2010 Only protein-protein interactions imex curation kong-2010-5 RNF220, an E3 ubiquitin ligase that targets Sin3B for ubiquitination. mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells fluorescence imaging fluorescence microscopy fluorescence imaging anti tag immunost anti tag immunostaining anti tag immunost Kong Q., Zeng W., Wu J., Hu W., Li C., Mao B. Apoptosis - Interactions involving proteins with a function related to apoptosis Ubiquitination assay. mao@mail.kiz.ac.cn Biochem. Biophys. Res. Commun. (0006-291X) 2010 Only protein-protein interactions imex curation ub2e1_mouse Ubiquitin-conjugating enzyme E2 E1 Ube2e1 Ubce5 Ubcm3 Ubiquitin-protein ligase E1 Ubiquitin carrier protein E1 UbcM3 (E3-independent) E2 ubiquitin-conjugating enzyme E1 E2 ubiquitin-conjugating enzyme E1 protein protein mouse Mus musculus Mouse MSDDDSRASTSSSSSSSSNQQTEKEGSTPKKKESKVSMSKNSKLLSTSAKRIQKELADITLDPPPNCSAGPKGDNIYEWRSTILGPPGSVYEGGVFFLDITFTPEYPFKPPKVTFRTRIYHCNINSQGVICLDILKDNWSPALTISKVLLSICSLLTDCNPADPLVGSIATQYMTNRAEHDRMARQWTKRYAT rn220_mouse E3 ubiquitin-protein ligase Rnf220 Rnf220 RING finger protein 220 RING-type E3 ubiquitin transferase Rnf220 protein protein mouse Mus musculus Mouse MDLHRAAFKMENSSYLPNPLASPALMVLASTAEASRDASIPCQQPRPFGVPVSVDKDVHIPFTNGSYTFASMYHRQGGVPGTFANRDFPPSLLHLHPQFAPPNLDCTPISMLNHSGVGAFRPFASTEDRESYQSAFTPAKRLKNCHDTESPHLRFSDADGKEYDFGTQLPSSSPGSLKVDDTGKKIFAVSGLISDRETSSSPEDRNDRCKKKAVALFDSQAPLCPICQVLLRPSELQEHMEQELEQLAQLPASKNSLLKDAMAPGTPKSLLLSASIKREGDSPTASPHSSATEDLHHSDRYQTFLRVRANRQTRLNARIGKMKRRKQDEGQREGSCMAEDDAVDIEHADSNRFEEYEWCGQKRIRATTLLEGGFRGSGFVMCSGKENPDSDADLDVDGDDTLEYGKPQYTEADVIPCTGEEPGEAKEREALRGAVLNGGPPSTRITPEFSKWASDEMPSTSNGEGSKQEAMQKTCKNSDIEKITEESAVTTFEALKARVRELERQLSRGDRYKCLICMDSYSMPLTSIQCWHVHCEECWLRTLGAKKLCPQCNTITAPGDLRRIYL ub2e3_mouse Ubiquitin-conjugating enzyme E2 E3 Ube2e3 Ubce4 Ubcm2 Ubiquitin-protein ligase E3 Ubiquitin carrier protein E3 Ubiquitin-conjugating enzyme E2-23 kDa UbcM2 E2 ubiquitin-conjugating enzyme E3 protein protein mouse Mus musculus Mouse MSSDRQRSDDESPSTSSGSSDADQRDPAAPEPEEQEERKPSATQQKKNTKLSSKTTAKLSTSAKRIQKELAEITLDPPPNCSAGPKGDNIYEWRSTILGPPGSVYEGGVFFLDITFSSDYPFKPPKVTFRTRIYHCNINSQGVICLDILKDNWSPALTISKVLLSICSLLTDCNPADPLVGSIATQYLTNRAEHDRIARQWTKRYAT sin3b_mouse Paired amphipathic helix protein Sin3b Transcriptional corepressor Sin3b Histone deacetylase complex subunit Sin3b Sin3b Kiaa0700 protein protein mouse Mus musculus Mouse MAHAGSGGSAGRGFGGSRWGRSGSGGHEKLPVHVEDALTYLDQVKIRFGSDPATYNGFLEIMKEFKSQSIDTPGVIRRVSQLFHEHPDLIVGFNAFLPLGYRIDIPKNGKLNIQSPLSSQDNSHSHGDCGEDFKQMSYKEDRGQVPLESDSVEFNNAISYVNKIKTRFLDHPEIYRSFLEILHTYQKEQLHTKGRPFRGMSEEEVFTEVANLFRGQEDLLSEFGQFLPEAKRSLFTGNGSCEMNSGQKNEEKSLEHNKKRSRPSLLRPVSAPAKKKMKLRGTKDLSIAAVGKYGTLQEFSFFDKVRRVLKSQEVYENFLRCIALFNQELVSGSELLQLVSPFLGKFPELFAQFKSFLGVKELSFAPPMSDRSGDGISREIDYASCKRIGSSYRALPKTYQQPKCSGRTAICKEVLNDTWVSFPSWSEDSTFVSSKKTPYEEQLHRCEDERFELDVVLETNLATIRVLESVQKKLSRMAPEDQEKLRLDDCLGGTSEVIQRRAIHRIYGDKAPEVIESLKRNPATAVPVVLKRLKAKEEEWREAQQGFNKIWREQYEKAYLKSLDHQAVNFKQNDTKALRSKSLLNEIESVYDEHQEQHSEGRSAPSSEPHLIFVYEDRQILEDAAALISYYVKRQPAIQKEDQGTIRQLLHRFLPSLFFSQQCPGTSDDSADERDRDRDSAEPERRRPTDEKPPADASPEPPKVLDDVYSLFFANNNWYFFLRLHQTLCARLLKIYRQAQKQLLEHRREQEREQLLCEGRREKAADPAMELRLKQPSEVELEEYYPAFLDMVRSLLEGSIDPTQYEDTLREMFTIHAYIGFTMDKLVQNIARQLHHLVSDDVCLKVVELYLNEQQRGAAGGNLSSRCVRAARETSYQWKAERCMADENCFKVMFLQRRGQVIMTIELLDTEEAQTEDPVEVQHLARYVEQYVGSEGASSSSTEGFLLKPVFLQRNLKKFRRWQCEQVRAMRGEAKSSWKRLMGVESACDVDCRFRLGTHKMVFIVNSEDYMYRRGTLCRAKQVQPLVLLRHHRHFEEWHGRWLEDNVTVAAAGLVQDWLMGEEEEDMVPCKTLCETAHVHGLPVTRYRVQYSRRPASP ub2d3_human Ubiquitin-conjugating enzyme E2 D3 UBC5C Ubiquitin-protein ligase D3 Ubiquitin carrier protein D3 Ubiquitin-conjugating enzyme E2-17 kDa 3 Ubiquitin-conjugating enzyme E2(17)KB 3 UBE2D3 UBCH5C (E3-independent) E2 ubiquitin-conjugating enzyme D3 E2 ubiquitin-conjugating enzyme D3 protein protein human Homo sapiens Human MALKRINKELSDLARDPPAQCSAGPVGDDMFHWQATIMGPNDSPYQGGVFFLTIHFPTDYPFKPPKVAFTTRIYHPNINSNGSICLDILRSQWSPALTISKVLLSICSLLCDPNPDDPLVPEIARIYKTDRDKYNRISREWTQKYAM uba1_rabit Ubiquitin-like modifier-activating enzyme 1 Ubiquitin-activating enzyme E1 UBA1 UBE1 protein protein rabit Oryctolagus cuniculus (Rabbit) Rabbit MSSSPLSKKRRVSGPDPKPGSNCSPAQSVLPQVPSAPTNGMAKNGSEADIDEGLYSRQLYVLGHEAMKRLQTSSVLVSGLRGLGVEIAKNIILGGVKAVTLHDQGTAQWADLSSQFYLREEDIGKNRAEVSQPRLAELNSYVPVTAYTGPLVEDFLSGFQVVVLTNSPLEDQLRVGEFCHSRGIKLVVADTRGLFGQLFCDFGEEMILTDSNGEQPLSTMVSMVTKDNPGVVTCLDEARHGFESGDFVSFSEVQGMTELNGNQPIEIKVLGPYTFSICDTSNFSDYIRGGIVSQVKVPKKISFKSLSASLAEPDFVMTDFAKFSRPAQLHIGFQALHKFCAQHSRPPRPRNEEDAAELVTLARAVNSKASSAVQQDSLDEDLIRNLAFVAAGDLAPINAFIGGLAAQEVMKACSGKFMPIMQWLYFDALECLPEDKESLTEDKCLPRQNRYDGQVAVFGSDLQEKLGRQKYFLVGAGAIGCELLKNFAMIGLGCGENGEIIVTDMDTIEKSNLNRQFLFRPWDVTKLKSDTAAAAVHQMNPHIRVTSHQNRVGPDTERIYDDDFFQTLDGVANALDNVDARMYMDRRCVYYRKPLLESGTLGTKGNVQVVIPFLTESYSSSQDPPEKSIPICTLKNFPNAIEHTLQWARDEFEGLFKQPAENVNQYLTDPKFVERTLRLAGTQPLEVLEAVQRSLVLQLPQSWADCVTWACHHWHTQYSNNIRQLLHNFPPDQLTSSGAPFWSGPKRCPHPLTFDVSNPLHLDYVMAAANLFAQTYGLAGSQDRAAVATLLQSVQVPEFTPKSGVKIHVSDQELQSANASVDDSRLEELKATLPSPDKLPGFKMYPIDFEKDDDSNFHMDFIVAASNLRAENYDIPPADRHKSKLIAGKIIPAIATTTAAVVGLVCLELYKVVQGHRHLDSYKNGFLNLALPFFGFSEPLAAPRHQYYNQEWTLWDRFEVQGLQPNGEEMTLKQFLDYFKTEHKLEITMLSQGVSMLYSFFMPAAKLKERLDQPMTEIVSRVSKRKLGRHVRALVLELCCNDESGEDVEVPYVRYTIR ubb_human Polyubiquitin-B UBB protein protein human Homo sapiens Human MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGC ube2e1-rnf220-1 Interaction between Rnf220 and Ube2e1 922990 n/a 922995 unspecified role unspecified role bait bait n-terminal 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 false n/a 922996 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 false physical association physical association false false false Fig. 1E ube2e3-rnf220-1 Interaction between Rnf220 and Ube2e3 922990 n/a 922997 unspecified role unspecified role bait bait n-terminal 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 false n/a 922996 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 false physical association physical association false false false Fig. 1E sin3b-rnf220-1 Interaction between Rnf220 and Sin3b 922990 n/a 922996 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 false n/a 922998 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 false pah1 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain range range false pah2 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range range range false pah3 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range range range false physical association physical association false false false Fig. 2A, 2D Exact coordinates of PAH domains not given in publication so have been inferred from the UniProtKB entry. sin3b-rnf220-2 Interaction between Rnf220 and Sin3b 922990 n/a 922996 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 922998 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 false physical association physical association false false false Figs. 2A, 2C A number of different domains of Rnf220 were seen as interacting with Sin3B. rnf220-rps27a-1 Auto-ubiquitination of Rnf220 922991 n/a 922999 unspecified role unspecified role ancillary ancillary region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 923000 unspecified role unspecified role ancillary ancillary rabit Oryctolagus cuniculus (Rabbit) Rabbit n/a 922996 putative self putative self putative self putative self ring domain Alters kinetics of interaction 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 923001 unspecified role unspecified role ancillary ancillary n-terminal 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 false ecoli Escherichia coli (strain K12) ubiquitination ubiquitination reaction ubiquitination false false false UBA1 purified; Ubiquitination of Rnf220 Fig. 1D sin3b-rnf220-3 Interaction between Rnf220 and Sin3b 922992 n/a 922996 unspecified role unspecified role bait bait n/a 922998 unspecified role unspecified role prey prey physical association physical association false false false sin3b-rnf220-4 Interaction between Rnf220 and Sin3b 922993 n/a 922996 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 922998 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 false in vitro In vitro physical association physical association false false false Figs. 2B sin3b-rnf220-5 Colocalisation of Rnf220 and Sin3b 922994 n/a 922998 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 922996 unspecified role unspecified role neutral component neutral component 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 false ring finger region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false colocalization colocalization false false false Figs. 5B