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/ UXT interacts with the transcriptional repressor protein EVI1 and suppresses cell transformation. UXT interacts with the transcriptional repressor protein EVI1 and suppresses cell transformation. FEBS J. (1742-464X) 2007 imex curation McGilvray R., Walker M., Bartholomew C. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer The range for the Rp domain of Evi-1 is stated in Palmer et al. (2001) J Biol Chem 276, 25830-25840. bartholomew@gcal.ac.uk Only protein-protein interactions 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 predetermined predetermined participant predetermined McGilvray R., Walker M., Bartholomew C. Apoptosis - Interactions involving proteins with a function related to apoptosis The range for the Rp domain of Evi-1 is stated in Palmer et al. (2001) J Biol Chem 276, 25830-25840. bartholomew@gcal.ac.uk Accepted 2007-OCT-4 by JYOTI. FEBS J. (1742-464X) 2007 Only protein-protein interactions imex curation UXT interacts with the transcriptional repressor protein EVI1 and suppresses cell transformation. UXT interacts with the transcriptional repressor protein EVI1 and suppresses cell transformation. FEBS J. (1742-464X) 2007 imex curation McGilvray R., Walker M., Bartholomew C. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer The range for the Rp domain of Evi-1 is stated in Palmer et al. (2001) J Biol Chem 276, 25830-25840. bartholomew@gcal.ac.uk Only protein-protein interactions 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 McGilvray R., Walker M., Bartholomew C. Apoptosis - Interactions involving proteins with a function related to apoptosis bartholomew@gcal.ac.uk Accepted 2007-OCT-4 by JYOTI. A human adult kidney cDNA library (Clontech) was used. FEBS J. (1742-464X) 2007 Only protein-protein interactions imex curation UXT interacts with the transcriptional repressor protein EVI1 and suppresses cell transformation. UXT interacts with the transcriptional repressor protein EVI1 and suppresses cell transformation. FEBS J. (1742-464X) 2007 imex curation McGilvray R., Walker M., Bartholomew C. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer The range for the Rp domain of Evi-1 is stated in Palmer et al. (2001) J Biol Chem 276, 25830-25840. bartholomew@gcal.ac.uk Only protein-protein interactions in vitro In vitro pull down pull down predetermined predetermined participant predetermined McGilvray R., Walker M., Bartholomew C. Apoptosis - Interactions involving proteins with a function related to apoptosis The range for the Rp domain of Evi-1 is stated in Palmer et al. (2001) J Biol Chem 276, 25830-25840. bartholomew@gcal.ac.uk Accepted 2007-OCT-14 by JYOTI. FEBS J. (1742-464X) 2007 Only protein-protein interactions imex curation UXT interacts with the transcriptional repressor protein EVI1 and suppresses cell transformation. UXT interacts with the transcriptional repressor protein EVI1 and suppresses cell transformation. FEBS J. (1742-464X) 2007 imex curation McGilvray R., Walker M., Bartholomew C. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer The range for the Rp domain of Evi-1 is stated in Palmer et al. (2001) J Biol Chem 276, 25830-25840. bartholomew@gcal.ac.uk Only protein-protein interactions human-bosc_23 Homo sapiens embryonic kidney cell line BOSC-23 bosc_23 Human embryonic kidney cell line BOSC-23 The BOSC-23 cell line are a derivative of the 293T cell line. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot McGilvray R., Walker M., Bartholomew C. Apoptosis - Interactions involving proteins with a function related to apoptosis bartholomew@gcal.ac.uk Accepted 2007-OCT-14 by JYOTI. FEBS J. (1742-464X) 2007 Only protein-protein interactions imex curation mecom_human Histone-lysine N-methyltransferase MECOM MECOM EVI1 Ecotropic virus integration site 1 protein homolog MDS1 Myelodysplasia syndrome 1 protein Myelodysplasia syndrome-associated protein 1 MDS1 and EVI1 complex locus protein PRDM3 protein protein human Homo sapiens Human MRSKGRARKLATNNECVYGNYPEIPLEEMPDADGVASTPSLNIQEPCSPATSSEAFTPKEGSPYKAPIYIPDDIPIPAEFELRESNMPGAGLGIWTKRKIEVGEKFGPYVGEQRSNLKDPSYGWEILDEFYNVKFCIDASQPDVGSWLKYIRFAGCYDQHNLVACQINDQIFYRVVADIAPGEELLLFMKSEDYPHETMAPDIHEERQYRCEDCDQLFESKAELADHQKFPCSTPHSAFSMVEEDFQQKLESENDLQEIHTIQECKECDQVFPDLQSLEKHMLSHTEEREYKCDQCPKAFNWKSNLIRHQMSHDSGKHYECENCAKVFTDPSNLQRHIRSQHVGARAHACPECGKTFATSSGLKQHKHIHSSVKPFICEVCHKSYTQFSNLCRHKRMHADCRTQIKCKDCGQMFSTTSSLNKHRRFCEGKNHFAAGGFFGQGISLPGTPAMDKTSMVNMSHANPGLADYFGANRHPAGLTFPTAPGFSFSFPGLFPSGLYHRPPLIPASSPVKGLSSTEQTNKSQSPLMTHPQILPATQDILKALSKHPSVGDNKPVELQPERSSEERPFEKISDQSESSDLDDVSTPSGSDLETTSGSDLESDIESDKEKFKENGKMFKDKVSPLQNLASINNKKEYSNHSIFSPSLEEQTAVSGAVNDSIKAIASIAEKYFGSTGLVGLQDKKVGALPYPSMFPLPFFPAFSQSMYPFPDRDLRSLPLKMEPQSPGEVKKLQKGSSESPFDLTTKRKDEKPLTPVPSKPPVTPATSQDQPLDLSMGSRSRASGTKLTEPRKNHVFGGKKGSNVESRPASDGSLQHARPTPFFMDPIYRVEKRKLTDPLEALKEKYLRPSPGFLFHPQMSAIENMAEKLESFSALKPEASELLQSVPSMFNFRAPPNALPENLLRKGKERYTCRYCGKIFPRSANLTRHLRTHTGEQPYRCKYCDRSFSISSNLQRHVRNIHNKEKPFKCHLCDRCFGQQTNLDRHLKKHENGNMSGTATSSPHSELESTGAILDDKEDAYFTEIRNFIGNSNHGSQSPRNVEERMNGSHFKDEKALVTSQNSDLLDDEEVEDEVLLDEEDEDNDITGKTGKEPVTSNLHEGNPEDDYEETSALEMSCKTSPVRYKEEEYKSGLSALDHIRHFTDSLKMRKMEDNQYSEAELSSFSTSHVPEELKQPLHRKSKSQAYAMMLSLSDKESLHSTSHSSSNVWHSMARAAAESSAIQSISHV DIP protein Q03112 original sequence version: 129 49E9B7CBCD422042 ctbp2_mouse C-terminal-binding protein 2 Ctbp2 protein protein mouse Mus musculus Mouse MALVDKHKVKRQRLDRICEGIRPQIMNGPLHPRPLVALLDGRDCTVEMPILKDLATVAFCDAQSTQEIHEKVLNEAVGAMMYHTITLTREDLEKFKALRVIVRIGSGYDNVDIKAAGELGIAVCNIPSAAVEETADSTVCHILNLYRRNTWLYQALREGTRVQSVEQIREVASGAARIRGETLGLIGFGRTGQAVAVRAKAFGFSVIFYDPYLQDGIERSLGVQRVYTLQDLLYQSDCVSLHCNLNEHNHHLINDFTIKQMRQGAFLVNAARGGLVDEKALAQALKEGRIRGAALDVHESEPFSFAQGPLKDAPNLICTPHTAWYSEQASLEMREAAATEIRRAITGRIPESLRNCVNKEFFVTSAPWSVIDQQAIHPELNGATYRYPPGIVGVAPGGLPPAMEGIIPGGIPVTHNLPTVAHPSQAPSPNQPTKHGDNREHPNEQ 5A81B9D75A9E493B uxt_human Protein UXT UXT HSPC024 Ubiquitously expressed transcript protein Androgen receptor trapped clone 27 protein protein protein human Homo sapiens Human MATPPKRRAVEATGEKVLRYETFISDVLQRDLRKVLDHRDKVYEQLAKYLQLRNVIERLQEAKHSELYMQVDLGCNFFVDTVVPDTSRIYVALGYGFFLELTLAEALKFIDRKSSLLTELSNSLTKDSMNIKAHIHMLLEGLRELQGLQNFPEKPHH 94CE14C462DEE308 evi1-ctbp2-1 1 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait region Rp domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 1 A, B, C, 5 D, E, F pKGIRp produces a GAL4 DNA-binding domain (DBD)Rp protein which interacts with mCtBP2 in yeast AH109 cells The Rp-deletion mutant lacking aa 706-707 retains ability to bind mCtBP2. evi1-uxt 2 5 unspecified role unspecified role bait bait region Rp domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role prey prey physical association physical association evi1-uxt-1 1 7 cofactor cofactor coenzyme bait bait 5 cofactor cofactor coenzyme prey prey region Rp domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association 1 A, B, C, 5 D, E, F uxt-uxt 1 7 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey physical association physical association 1 A, B, C uxt-evi1 3 7 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 ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction 3 B GST pulldown assays were performed with combinations of bacterially expressed and 35S-methionine-labelled proteins and the results are shown in Fig. 3. These confirm that UXT binds Rp evi1-uxt-2 3 5 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 ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction 3 B GST pulldown assays were performed with combinations of bacterially expressed and 35S-methionine-labelled proteins and the results are shown in Fig. 3. These confirm that UXT binds Rp evi1-uxt-3 4 5 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 7 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 3 C Immunoprecipitation of cell extracts and western blot analysis with a-myc reveals EVI1myc in cells transfected with pcDNA3EVI1myc, as expected (Fig. 3; IP a-9E10). In addition, western blot analysis of a-myc-immunoprecipitated cell extracts with a-HA shows HAUXT only in those extracts that also contain EVI1myc, confirming that these two proteins form a complex. Q03112; Q9UBK9 transfection.