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/ The E3 ligase RING1 targets p53 for degradation and promotes cancer cell proliferation and survival. The E3 ligase RING1 targets p53 for degradation and promotes cancer cell proliferation and survival. Cancer research 2017 imex curation Shen J., Li P., Shao X., Yang Y., Liu XJ., Feng M., Yu Q., Hu R., Wang Z. Cancer - Interactions investigated in the context of cancer The authors have stated that all constructs used were of human origin. wangzhen@imb.pumc.edu.cn,coryhu@sibcb.ac.cn 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-hct116 Human colon carcinoma HCT116 cells hct116 Human colon carcinoma line HCT116 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Shen J., Li P., Shao X., Yang Y., Liu XJ., Feng M., Yu Q., Hu R., Wang Z. Cancer - Interactions investigated in the context of cancer The authors have stated that all constructs used were of human origin. wangzhen@imb.pumc.edu.cn,coryhu@sibcb.ac.cn Accepted 2018-FEB-01 AT 12:30 GMT AT 12:30 GMT by LIVIAP Cancer research 2017 imex curation Just had a word with Pablo:- he agrees that there are only the "enzyme" and target that are participants - the rest are ancillarys, so in effect, do not count! There are lots of other examples of interactions like this in the Database (if you can access it!) e.g. EBI-6664613 The E3 ligase RING1 targets p53 for degradation and promotes cancer cell proliferation and survival. The E3 ligase RING1 targets p53 for degradation and promotes cancer cell proliferation and survival. Cancer research 2017 imex curation Shen J., Li P., Shao X., Yang Y., Liu XJ., Feng M., Yu Q., Hu R., Wang Z. Cancer - Interactions investigated in the context of cancer The authors have stated that all constructs used were of human origin. wangzhen@imb.pumc.edu.cn,coryhu@sibcb.ac.cn 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Shen J., Li P., Shao X., Yang Y., Liu XJ., Feng M., Yu Q., Hu R., Wang Z. Cancer - Interactions investigated in the context of cancer The authors have stated that all constructs used were of human origin. wangzhen@imb.pumc.edu.cn,coryhu@sibcb.ac.cn Accepted 2018-FEB-01 AT 12:30 GMT AT 12:30 GMT by LIVIAP Cancer research 2017 imex curation Just had a word with Pablo:- he agrees that there are only the "enzyme" and target that are participants - the rest are ancillarys, so in effect, do not count! There are lots of other examples of interactions like this in the Database (if you can access it!) e.g. EBI-6664613 The E3 ligase RING1 targets p53 for degradation and promotes cancer cell proliferation and survival. The E3 ligase RING1 targets p53 for degradation and promotes cancer cell proliferation and survival. Cancer research 2017 imex curation Shen J., Li P., Shao X., Yang Y., Liu XJ., Feng M., Yu Q., Hu R., Wang Z. Cancer - Interactions investigated in the context of cancer The authors have stated that all constructs used were of human origin. wangzhen@imb.pumc.edu.cn,coryhu@sibcb.ac.cn 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Shen J., Li P., Shao X., Yang Y., Liu XJ., Feng M., Yu Q., Hu R., Wang Z. Cancer - Interactions investigated in the context of cancer The authors have stated that all constructs used were of human origin. wangzhen@imb.pumc.edu.cn,coryhu@sibcb.ac.cn Accepted 2018-FEB-01 AT 12:30 GMT AT 12:30 GMT by LIVIAP Cancer research 2017 imex curation Just had a word with Pablo:- he agrees that there are only the "enzyme" and target that are participants - the rest are ancillarys, so in effect, do not count! There are lots of other examples of interactions like this in the Database (if you can access it!) e.g. EBI-6664613 The E3 ligase RING1 targets p53 for degradation and promotes cancer cell proliferation and survival. The E3 ligase RING1 targets p53 for degradation and promotes cancer cell proliferation and survival. Cancer research 2017 imex curation Shen J., Li P., Shao X., Yang Y., Liu XJ., Feng M., Yu Q., Hu R., Wang Z. Cancer - Interactions investigated in the context of cancer The authors have stated that all constructs used were of human origin. wangzhen@imb.pumc.edu.cn,coryhu@sibcb.ac.cn 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro ubiquitinase assay ubiquitinase assay ubiquitinase assay predetermined predetermined participant predetermined Shen J., Li P., Shao X., Yang Y., Liu XJ., Feng M., Yu Q., Hu R., Wang Z. Cancer - Interactions investigated in the context of cancer the method in the M+M section states HA tag was used - but NO HA tag on any constructs used - E1 from Ref 23. Method in Ref 23 cannot have been used here either?? So no idea of expression systems/prep for these proteins. The authors have stated that all constructs used were of human origin. wangzhen@imb.pumc.edu.cn,coryhu@sibcb.ac.cn Accepted 2018-FEB-01 AT 12:30 GMT AT 12:30 GMT by LIVIAP Cancer research 2017 imex curation Just had a word with Pablo:- he agrees that there are only the "enzyme" and target that are participants - the rest are ancillarys, so in effect, do not count! There are lots of other examples of interactions like this in the Database (if you can access it!) e.g. EBI-6664613 The E3 ligase RING1 targets p53 for degradation and promotes cancer cell proliferation and survival. The E3 ligase RING1 targets p53 for degradation and promotes cancer cell proliferation and survival. Cancer research 2017 imex curation Shen J., Li P., Shao X., Yang Y., Liu XJ., Feng M., Yu Q., Hu R., Wang Z. Cancer - Interactions investigated in the context of cancer The authors have stated that all constructs used were of human origin. wangzhen@imb.pumc.edu.cn,coryhu@sibcb.ac.cn 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-hct116 Human colon carcinoma HCT116 cells hct116 Human colon carcinoma line HCT116 fluorescence imaging fluorescence microscopy fluorescence imaging immunocytochemistry immunocytochemistry Shen J., Li P., Shao X., Yang Y., Liu XJ., Feng M., Yu Q., Hu R., Wang Z. Cancer - Interactions investigated in the context of cancer The authors have stated that all constructs used were of human origin. wangzhen@imb.pumc.edu.cn,coryhu@sibcb.ac.cn Accepted 2018-FEB-01 AT 12:30 GMT AT 12:30 GMT by LIVIAP Cancer research 2017 imex curation Just had a word with Pablo:- he agrees that there are only the "enzyme" and target that are participants - the rest are ancillarys, so in effect, do not count! There are lots of other examples of interactions like this in the Database (if you can access it!) e.g. EBI-6664613 The E3 ligase RING1 targets p53 for degradation and promotes cancer cell proliferation and survival. The E3 ligase RING1 targets p53 for degradation and promotes cancer cell proliferation and survival. Cancer research 2017 imex curation Shen J., Li P., Shao X., Yang Y., Liu XJ., Feng M., Yu Q., Hu R., Wang Z. Cancer - Interactions investigated in the context of cancer The authors have stated that all constructs used were of human origin. wangzhen@imb.pumc.edu.cn,coryhu@sibcb.ac.cn 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). fluorescence imaging fluorescence microscopy fluorescence imaging immunocytochemistry immunocytochemistry Shen J., Li P., Shao X., Yang Y., Liu XJ., Feng M., Yu Q., Hu R., Wang Z. Cancer - Interactions investigated in the context of cancer The authors have stated that all constructs used were of human origin. wangzhen@imb.pumc.edu.cn,coryhu@sibcb.ac.cn Accepted 2018-FEB-01 AT 12:30 GMT AT 12:30 GMT by LIVIAP Cancer research 2017 imex curation Just had a word with Pablo:- he agrees that there are only the "enzyme" and target that are participants - the rest are ancillarys, so in effect, do not count! There are lots of other examples of interactions like this in the Database (if you can access it!) e.g. EBI-6664613 ub2e1_human Ubiquitin-conjugating enzyme E2 E1 UBE2E1 UBCH6 Ubiquitin-protein ligase E1 Ubiquitin carrier protein E1 UbcH6 (E3-independent) E2 ubiquitin-conjugating enzyme E1 E2 ubiquitin-conjugating enzyme E1 protein protein human Homo sapiens Human MSDDDSRASTSSSSSSSSNQQTEKETNTPKKKESKVSMSKNSKLLSTSAKRIQKELADITLDPPPNCSAGPKGDNIYEWRSTILGPPGSVYEGGVFFLDITFTPEYPFKPPKVTFRTRIYHCNINSQGVICLDILKDNWSPALTISKVLLSICSLLTDCNPADPLVGSIATQYMTNRAEHDRMARQWTKRYAT 2FBC50BE2A6A0008 uba1_human Ubiquitin-like modifier-activating enzyme 1 Ubiquitin-activating enzyme E1 Protein A1S9 UBA1 A1S9T UBE1 protein protein human Homo sapiens Human MSSSPLSKKRRVSGPDPKPGSNCSPAQSVLSEVPSVPTNGMAKNGSEADIDEGLYSRQLYVLGHEAMKRLQTSSVLVSGLRGLGVEIAKNIILGGVKAVTLHDQGTAQWADLSSQFYLREEDIGKNRAEVSQPRLAELNSYVPVTAYTGPLVEDFLSGFQVVVLTNTPLEDQLRVGEFCHNRGIKLVVADTRGLFGQLFCDFGEEMILTDSNGEQPLSAMVSMVTKDNPGVVTCLDEARHGFESGDFVSFSEVQGMVELNGNQPMEIKVLGPYTFSICDTSNFSDYIRGGIVSQVKVPKKISFKSLVASLAEPDFVVTDFAKFSRPAQLHIGFQALHQFCAQHGRPPRPRNEEDAAELVALAQAVNARALPAVQQNNLDEDLIRKLAYVAAGDLAPINAFIGGLAAQEVMKACSGKFMPIMQWLYFDALECLPEDKEVLTEDKCLQRQNRYDGQVAVFGSDLQEKLGKQKYFLVGAGAIGCELLKNFAMIGLGCGEGGEIIVTDMDTIEKSNLNRQFLFRPWDVTKLKSDTAAAAVRQMNPHIRVTSHQNRVGPDTERIYDDDFFQNLDGVANALDNVDARMYMDRRCVYYRKPLLESGTLGTKGNVQVVIPFLTESYSSSQDPPEKSIPICTLKNFPNAIEHTLQWARDEFEGLFKQPAENVNQYLTDPKFVERTLRLAGTQPLEVLEAVQRSLVLQRPQTWADCVTWACHHWHTQYSNNIRQLLHNFPPDQLTSSGAPFWSGPKRCPHPLTFDVNNPLHLDYVMAAANLFAQTYGLTGSQDRAAVATFLQSVQVPEFTPKSGVKIHVSDQELQSANASVDDSRLEELKATLPSPDKLPGFKMYPIDFEKDDDSNFHMDFIVAASNLRAENYDIPSADRHKSKLIAGKIIPAIATTTAAVVGLVCLELYKVVQGHRQLDSYKNGFLNLALPFFGFSEPLAAPRHQYYNQEWTLWDRFEVQGLQPNGEEMTLKQFLDYFKTEHKLEITMLSQGVSMLYSFFMPAAKLKERLDQPMTEIVSRVSKRKLGRHVRALVLELCCNDESGEDVEVPYVRYTIR DIP protein P22314 original sequence version: 157 DIP protein P22314 original sequence version: 152 4B413AAA75FAA562 p53_human Cellular tumor antigen p53 TP53 P53 Tumor suppressor p53 Phosphoprotein p53 Antigen NY-CO-13 protein protein human Homo sapiens Human MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLSPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPPEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMFRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD DIP protein P04637 original sequence version: 121 DIP protein P04637 original sequence version: 115 DIP protein P04637-1 original sequence version: 224 DIP protein P04637 original sequence version: 117 DIP protein P04637-1 original sequence version: 226 DIP protein P04637 original sequence version: 226 DIP protein P04637 original sequence version: 212 DIP protein P04637 original sequence version: 220 DIP protein P04637-1 original sequence version: 220 DIP protein P04637-1 original sequence version: 145 DIP protein P04637 original sequence version: 146 DIP protein P04637 original sequence version: 145 DIP protein P04637 original sequence version: 166 DIP protein P04637-1 original sequence version: 166 DIP protein P04637 original sequence version: 169 DIP protein P04637 original sequence version: 177 DIP protein P04637 original sequence version: 175 DIP protein P04637-1 original sequence version: 175 DIP protein P04637-1 original sequence version: 172 DIP protein P04637-1 original sequence version: 169 DIP protein P04637 original sequence version: 170 DIP protein P04637 original sequence version: 172 DIP protein P04637-1 original sequence version: 194 DIP protein P04637 original sequence version: 196 DIP protein P04637 original sequence version: 185 DIP protein P04637-1 original sequence version: 191 DIP protein P04637 original sequence version: 191 DIP protein P04637 original sequence version: 180 DIP protein P04637-1 original sequence version: 197 DIP protein P04637 original sequence version: 183 DIP protein P04637-1 original sequence version: 183 DIP protein P04637-1 original sequence version: 185 DIP protein P04637 original sequence version: 199 DIP protein P04637 original sequence version: 201 DIP protein P04637 original sequence version: 189 DIP protein P04637 original sequence version: 235 DIP protein P04637 original sequence version: 230 DIP protein P04637 original sequence version: 234 DIP protein P04637-1 original sequence version: 234 DIP protein P04637-1 original sequence version: 235 DIP protein P04637 original sequence version: 239 DIP protein P04637-1 original sequence version: 212 DIP protein P04637-1 original sequence version: 213 DIP protein P04637 original sequence version: 213 DIP protein P04637 original sequence version: 206 DIP protein P04637-1 original sequence version: 188 DIP protein P04637-1 original sequence version: 228 DIP protein P04637 original sequence version: 208 DIP protein P04637 original sequence version: 240 DIP protein P04637-1 original sequence version: 240 DIP protein P04637 original sequence version: 204 DIP protein P04637-1 original sequence version: 204 DIP protein P04637-1 original sequence version: 209 DIP protein P04637-1 original sequence version: 203 DIP protein P04637 original sequence version: 221 DIP protein P04637-1 original sequence version: 152 DIP protein P04637 original sequence version: 217 DIP protein P04637-1 original sequence version: 140 DIP protein P04637 original sequence version: 215 DIP protein P04637 original sequence version: 214 DIP protein P04637-1 original sequence version: 182 DIP protein P04637 original sequence version: 182 DIP protein P04637 original sequence version: 188 DIP protein P04637 original sequence version: 192 DIP protein P04637-1 original sequence version: 192 DIP protein P04637 original sequence version: 218 DIP protein P04637-1 original sequence version: 218 AD5C149FD8106131 ring1_human E3 ubiquitin-protein ligase RING1 RING finger protein 1 Polycomb complex protein RING1 RING1 RNF1 Really interesting new gene 1 protein RING-type E3 ubiquitin transferase RING1 protein protein human Homo sapiens Human MTTPANAQNASKTWELSLYELHRTPQEAIMDGTEIAVSPRSLHSELMCPICLDMLKNTMTTKECLHRFCSDCIVTALRSGNKECPTCRKKLVSKRSLRPDPNFDALISKIYPSREEYEAHQDRVLIRLSRLHNQQALSSSIEEGLRMQAMHRAQRVRRPIPGSDQTTTMSGGEGEPGEGEGDGEDVSSDSAPDSAPGPAPKRPRGGGAGGSSVGTGGGGTGGVGGGAGSEDSGDRGGTLGGGTLGPPSPPGAPSPPEPGGEIELVFRPHPLLVEKGEYCQTRYVKTTGNATVDHLSKYLALRIALERRQQQEAGEPGGPGGGASDTGGPDGCGGEGGGAGGGDGPEEPALPSLEGVSEKQYTIYIAPGGGAFTTLNGSLTLELVNEKFWKVSRPLELCYAPTKDPK DIP protein Q06587 original sequence version: 103 DIP protein Q06587 original sequence version: 147 DIP protein Q06587 original sequence version: 135 6959787479DE9DAB ring1-tp53-1 3 9 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 ring_domain required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain Q06587:p.Ile50Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain I A Wed May 02 19:09:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile50ala' Tue Jun 30 18:07:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile50Ala' physical association physical association Figure 3A, 4A, Supplementary Figure 2D, 3C Supplementary Figure 2D;this interaction has been assumed to take place in 293T cells, as referred to in relation to Figure 4C, but is not explicitly stated. This interaction occurs in RNF2-/- cells. ring1-tp53-2 6 9 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell 10 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization Figure 3B ring1-tp53-3 5 9 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell 10 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization Figure 3B tp53-ring1-1 2 10 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. 9 unspecified role unspecified role bait bait purified purified n_terminus 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 ecoli Escherichia coli (strain K12) physical association physical association Figure 3C ring1-tp53-4 4 9 enzyme target enzyme target substrate substrate neutral component neutral component purified purified 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 region ubiquitinylated lysine ubiquitinylated lysine undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined resulting-ptm resulting-ptm resulting ptm resulting-ptm 10 enzyme enzyme neutral component neutral component purified purified 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 region mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 7 ancillary ancillary neutral component neutral component purified purified 8 ancillary ancillary neutral component neutral component purified purified ubiquitination ubiquitination reaction ubiquitination Figure 3E ring1-tp53-5 1 9 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Supplementary figure 2A tp53-ring1-2 2 10 unspecified role unspecified role bait bait purified purified region mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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 ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus 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 ecoli Escherichia coli (strain K12) physical association physical association Supplementary Figure 2B