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/ Stabilization of snail by NF-kappaB is required for inflammation-induced cell migration and invasion. Stabilization of snail by NF-kappaB is required for inflammation-induced cell migration and invasion. Cancer Cell (1535-6108) 2009 imex curation Wu Y., Deng J., Rychahou PG., Qiu S., Evers BM., Zhou BP. Cancer - Interactions investigated in the context of cancer Proteasome inhibitor MG-132 Details of constructs provided by authors. peter.zhou@uky.edu,bpzhou@utmb.edu 25-Jun-2010: Contacted by JYOTI. orchard Only protein-protein interactions 7 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 Wu Y., Deng J., Rychahou PG., Qiu S., Evers BM., Zhou BP. Cancer - Interactions investigated in the context of cancer Proteasome inhibitor MG-132 Details of constructs provided by authors. peter.zhou@uky.edu peter.zhou@uky.edu,bpzhou@utmb.edu Accepted 2010-APR-15 by ORCHARD. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 7 imex curation Stabilization of snail by NF-kappaB is required for inflammation-induced cell migration and invasion. Stabilization of snail by NF-kappaB is required for inflammation-induced cell migration and invasion. Cancer Cell (1535-6108) 2009 imex curation Wu Y., Deng J., Rychahou PG., Qiu S., Evers BM., Zhou BP. Cancer - Interactions investigated in the context of cancer Proteasome inhibitor MG-132 Details of constructs provided by authors. peter.zhou@uky.edu,bpzhou@utmb.edu 25-Jun-2010: Contacted by JYOTI. orchard Only protein-protein interactions 7 human-pc3 Human PC3 Caucasian prostate adenocarcinoma pc3 Human Caucasian prostate adenocarcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Wu Y., Deng J., Rychahou PG., Qiu S., Evers BM., Zhou BP. Cancer - Interactions investigated in the context of cancer Proteasome inhibitor MG-132 Details of constructs provided by authors. peter.zhou@uky.edu peter.zhou@uky.edu,bpzhou@utmb.edu Accepted 2010-APR-15 by ORCHARD. 25-Jun-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 7 imex curation Stabilization of snail by NF-kappaB is required for inflammation-induced cell migration and invasion. Stabilization of snail by NF-kappaB is required for inflammation-induced cell migration and invasion. Cancer Cell (1535-6108) 2009 imex curation Wu Y., Deng J., Rychahou PG., Qiu S., Evers BM., Zhou BP. Cancer - Interactions investigated in the context of cancer Proteasome inhibitor MG-132 Details of constructs provided by authors. peter.zhou@uky.edu,bpzhou@utmb.edu 25-Jun-2010: Contacted by JYOTI. orchard Only protein-protein interactions 7 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. fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Wu Y., Deng J., Rychahou PG., Qiu S., Evers BM., Zhou BP. Cancer - Interactions investigated in the context of cancer Proteasome inhibitor MG-132 Details of constructs provided by authors. peter.zhou@uky.edu peter.zhou@uky.edu,bpzhou@utmb.edu Accepted 2010-APR-15 by ORCHARD. 25-Jun-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 7 imex curation Stabilization of snail by NF-kappaB is required for inflammation-induced cell migration and invasion. Stabilization of snail by NF-kappaB is required for inflammation-induced cell migration and invasion. Cancer Cell (1535-6108) 2009 imex curation Wu Y., Deng J., Rychahou PG., Qiu S., Evers BM., Zhou BP. Cancer - Interactions investigated in the context of cancer Proteasome inhibitor MG-132 Details of constructs provided by authors. peter.zhou@uky.edu,bpzhou@utmb.edu 25-Jun-2010: Contacted by JYOTI. orchard Only protein-protein interactions 7 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 Wu Y., Deng J., Rychahou PG., Qiu S., Evers BM., Zhou BP. Cancer - Interactions investigated in the context of cancer Proteasome inhibitor MG-132 Details of constructs provided by authors. peter.zhou@uky.edu peter.zhou@uky.edu,bpzhou@utmb.edu Accepted 2010-APR-15 by ORCHARD. 25-Jun-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 7 imex curation snai1_human Zinc finger protein SNAI1 Protein snail homolog 1 SNAI1 SNAH protein protein human Homo sapiens Human MPRSFLVRKPSDPNRKPNYSELQDSNPEFTFQQPYDQAHLLAAIPPPEILNPTASLPMLIWDSVLAPQAQPIAWASLRLQESPRVAELTSLSDEDSGKGSQPPSPPSPAPSSFSSTSVSSLEAEAYAAFPGLGQVPKQLAQLSEAKDLQARKAFNCKYCNKEYLSLGALKMHIRSHTLPCVCGTCGKAFSRPWLLQGHVRTHTGEKPFSCPHCSRAFADRSNLRAHLQTHSDVKKYQCQACARTFSRMSLLHKHQESGCSGCPR DIP protein O95863 original sequence version: 109 DIP protein O95863 original sequence version: 141 70E298C9BB154115 gsk3b_human Glycogen synthase kinase-3 beta GSK3B Serine/threonine-protein kinase GSK3B protein protein human Homo sapiens Human MSGRPRTTSFAESCKPVQQPSAFGSMKVSRDKDGSKVTTVVATPGQGPDRPQEVSYTDTKVIGNGSFGVVYQAKLCDSGELVAIKKVLQDKRFKNRELQIMRKLDHCNIVRLRYFFYSSGEKKDEVYLNLVLDYVPETVYRVARHYSRAKQTLPVIYVKLYMYQLFRSLAYIHSFGICHRDIKPQNLLLDPDTAVLKLCDFGSAKQLVRGEPNVSYICSRYYRAPELIFGATDYTSSIDVWSAGCVLAELLLGQPIFPGDSGVDQLVEIIKVLGTPTREQIREMNPNYTEFKFPQIKAHPWTKVFRPRTPPEAIALCSRLLEYTPTARLTPLEACAHSFFDELRDPNVKLPNGRDTPALFNFTTQELSSNPPLATILIPPHARIQAAASTPTNATAASDANTGDRGQTNNAASASASNST DIP protein P49841 original sequence version: 109 DIP protein P49841 original sequence version: 136 DIP protein P49841 original sequence version: 143 DIP protein P49841 original sequence version: 141 DIP protein P49841 original sequence version: 196 DIP protein P49841 original sequence version: 159 DIP protein P49841 original sequence version: 151 4ACC24D00CDBB9C3 tf65_human Transcription factor p65 Nuclear factor NF-kappa-B p65 subunit RELA NFKB3 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 protein protein human Homo sapiens Human MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGSIPGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAELCPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQVTVRDPSGRPLRLPPVLSHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQKEDIEVYFTGPGWEARGSFSQADVHRQVAIVFRTPPYADPSLQAPVRVSMQLRRPSDRELSEPMEFQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISS DIP protein Q04206 original sequence version: 170 DIP protein Q04206 original sequence version: 173 DIP protein Q04206 original sequence version: 99 DIP protein Q04206-1 original sequence version: 99 DIP protein Q04206 original sequence version: 168 DIP protein Q04206 original sequence version: 128 DIP protein Q04206 original sequence version: 131 DIP protein Q04206 original sequence version: 153 DIP protein Q04206 original sequence version: 185 DIP protein Q04206 original sequence version: 181 DIP protein Q04206 original sequence version: 183 8147A6AF9F2445C6 csn2_human COP9 signalosome complex subunit 2 JAB1-containing signalosome subunit 2 Thyroid receptor-interacting protein 15 Alien homolog COPS2 CSN2 TRIP15 protein protein human Homo sapiens Human MSDMEDDFMCDDEEDYDLEYSEDSNSEPNVDLENQYYNSKALKEDDPKAALSSFQKVLELEGEKGEWGFKALKQMIKINFKLTNFPEMMNRYKQLLTYIRSAVTRNYSEKSINSILDYISTSKQMDLLQEFYETTLEALKDAKNDRLWFKTNTKLGKLYLEREEYGKLQKILRQLHQSCQTDDGEDDLKKGTQLLEIYALEIQMYTAQKNNKKLKALYEQSLHIKSAIPHPLIMGVIRECGGKMHLREGEFEKAHTDFFEAFKNYDESGSPRRTTCLKYLVLANMLMKSGINPFDSQEAKPYKNDPEILAMTNLVSAYQNNDITEFEKILKTNHSNIMDDPFIREHIEELLRNIRTQVLIKLIKPYTRIHIPFISKELNIDVADVESLLVQCILDNTIHGRIDQVNQLLELDHQKRGGARYTALDKWTNQLNSLNQAVVSKLA DIP protein P61201 original sequence version: 67 DIP protein P61201-1 original sequence version: 106 DIP protein P61201 original sequence version: 122 DIP protein P61201-1 original sequence version: 76 1DB6FA774C13BC59 fbw1a_human F-box/WD repeat-containing protein 1A BTRC BTRCP FBW1A FBXW1A F-box and WD repeats protein beta-TrCP E3RSIkappaB pIkappaBalpha-E3 receptor subunit Epididymis tissue protein Li 2a protein protein human Homo sapiens Human MDPAEAVLQEKALKFMCSMPRSLWLGCSSLADSMPSLRCLYNPGTGALTAFQNSSEREDCNNGEPPRKIIPEKNSLRQTYNSCARLCLNQETVCLASTAMKTENCVAKTKLANGTSSMIVPKQRKLSASYEKEKELCVKYFEQWSESDQVEFVEHLISQMCHYQHGHINSYLKPMLQRDFITALPARGLDHIAENILSYLDAKSLCAAELVCKEWYRVTSDGMLWKKLIERMVRTDSLWRGLAERRGWGQYLFKNKPPDGNAPPNSFYRALYPKIIQDIETIESNWRCGRHSLQRIHCRSETSKGVYCLQYDDQKIVSGLRDNTIKIWDKNTLECKRILTGHTGSVLCLQYDERVIITGSSDSTVRVWDVNTGEMLNTLIHHCEAVLHLRFNNGMMVTCSKDRSIAVWDMASPTDITLRRVLVGHRAAVNVVDFDDKYIVSASGDRTIKVWNTSTCEFVRTLNGHKRGIACLQYRDRLVVSGSSDNTIRLWDIECGACLRVLEGHEELVRCIRFDNKRIVSGAYDGKIKVWDLVAALDPRAPAGTLCLRTLVEHSGRVFRLQFDEFQIVSSSHDDTILIWDFLNDPAAQAEPPRSPSRTYTYISR DIP protein Q9Y297 original sequence version: 142 DIP protein Q9Y297 original sequence version: 153 DIP protein Q9Y297 original sequence version: 144 DIP protein Q9Y297 original sequence version: 145 DIP protein Q9Y297 original sequence version: 106 DIP protein Q9Y297 original sequence version: 82 DIP protein Q9Y297 original sequence version: 111 DIP protein Q9Y297 original sequence version: 125 4C67F3B7E400FD37 snail-gsk 1 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association Fig 6D. When endogenous GSK-3beta was immunoprecipitated, the associated endogenous Snail was significantly reduced in cells treated with TNFalpha, compared with that in cells treated with MG-132. TNFalpha snail-gsk-1 1 5 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association Fig 6E. Similarly, when endogenous Snail was pulled down, the bound endogenous GSK-3beta was lost in cells treated with TNFalpha. TNFalpha snail-gsk-2 2 5 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association Fig 6E. Similarly, when endogenous Snail was pulled down, the bound endogenous GSK-3beta was lost in cells treated with TNFalpha. TNFalpha p65-snail 3 5 unspecified role unspecified role neutral component neutral component 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 7 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig 3C. The expression of p65 dramatically enhanced Snail protein levels as the expression of p65 was colocalized with GFP-Snail in the nucleus. csn2-snail 3 8 unspecified role unspecified role neutral component neutral component 5 unspecified role unspecified role neutral component neutral component 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 colocalization colocalization Fig 5D. When CSN2 was coexpressed with GFP-Snail, it enhanced the intensity of GFP-Snail in the nucleus. snail-gsk-3 4 6 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 5 unspecified role unspecified role bait bait physical association physical association Fig 6C. The association of Snail with GSK-3beta was diminished in cells treated with TNFalpha or CSN2, compared with cells treated with MG-132. snail-btrc-1 4 9 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait region xlnk-n6lys-1gly N6-glycyl-L-lysine (2S)-2-amino-6-[(aminoacetyl)amino]hexanoic acid N6-(glycyl)-L-lysine N6-glycyllysine XLNK-N6Lys-1Gly CROSSLNK Glycyl lysine isopeptide (Gly-Lys) (interchain with K-...) CROSSLNK Glycyl lysine isopeptide (Lys-Gly) (interchain with G-...) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig 6B. The association of Snail with its E3 ligase, beta-Trcp, was abolished in cells treated with TNFa or expression with CSN2. Proteosome inhibitor, MG-132