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/ SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. Mol. Cell (1097-2765) 2006 Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov Accepted 2007-FEB-14 by DAVET. Human Hela cDNA library 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library Mol. Cell (1097-2765) 2006 SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. Mol. Cell (1097-2765) 2006 Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov Accepted 2007-FEB-14 by DAVET. Human Hela cDNA library 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library Mol. Cell (1097-2765) 2006 SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. Mol. Cell (1097-2765) 2006 Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer I searched for the CAD and cyclin B1 genes in human VEGA and used the sequences there to search for where the primers matched. Then I took the sequences determined from there as demarked by the primers and used NCBI nucleotide-nucleotide BLAST to find the accession number of the sequence containing the relevant sequence and this also showed the relevant ranges. fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov Accepted 2007-FEB-14 by DAVET. Human Hela cDNA library 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library Mol. Cell (1097-2765) 2006 SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. Mol. Cell (1097-2765) 2006 Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov Accepted 2007-FEB-14 by DAVET. Human Hela cDNA library 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library Mol. Cell (1097-2765) 2006 SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. Mol. Cell (1097-2765) 2006 Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov Accepted 2007-FEB-14 by DAVET. Human Hela cDNA library 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library Mol. Cell (1097-2765) 2006 SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. Mol. Cell (1097-2765) 2006 Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library 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 Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov Accepted 2007-FEB-14 by DAVET. Human Hela cDNA library 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library Mol. Cell (1097-2765) 2006 SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. SNIP1 is a candidate modifier of the transcriptional activity of c-Myc on E box-dependent target genes. Mol. Cell (1097-2765) 2006 Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Fujii M., Lyakh LA., Bracken CP., Fukuoka J., Hayakawa M., Tsukiyama T., Soll SJ., Harris M., Rocha S., Roche KC., Tominaga S., Jen J., Perkins ND., Lechleider RJ., Roberts AB. Cancer - Interactions investigated in the context of cancer fujiim@jichi.ac.jp,lechleiderr@mail.nih.gov Accepted 2007-FEB-14 by DAVET. Human Hela cDNA library 02-MAR-2006: Contacted by JYOTI. Human Hela cDNA library Mol. Cell (1097-2765) 2006 snip1_human Smad nuclear-interacting protein 1 SNIP1 FHA domain-containing protein SNIP1 protein protein human Homo sapiens Human MKAVKSERERGSRRRHRDGDVVLPAGVVVKQERLSPEVAPPAHRRPDHSGGSPSPPTSEPARSGHRGNRARGVSRSPPKKKNKASGRRSKSPRSKRNRSPHHSTVKVKQEREDHPRRGREDRQHREPSEQEHRRARNSDRDRHRGHSHQRRTSNERPGSGQGQGRDRDTQNLQAQEEEREFYNARRREHRQRNDVGGGGSESQELVPRPGGNNKEKEVPAKEKPSFELSGALLEDTNTFRGVVIKYSEPPEARIPKKRWRLYPFKNDEVLPVMYIHRQSAYLLGRHRRIADIPIDHPSCSKQHAVFQYRLVEYTRADGTVGRRVKPYIIDLGSGNGTFLNNKRIEPQRYYELKEKDVLKFGFSSREYVLLHESSDTSEIDRKDDEDEEEEEEVSDS B183F83EC3184676 max_human Protein max MAX BHLHD4 Class D basic helix-loop-helix protein 4 Myc-associated factor X protein protein human Homo sapiens Human MSDNDDIEVESDEEQPRFQSAADKRAHHNALERKRRDHIKDSFHSLRDSVPSLQGEKASRAQILDKATEYIQYMRRKNHTHQQDIDDLKRQNALLEQQVRALEKARSSAQLQTNYPSSDNSLYTNAKGSTISAFDGGSDSSSESEPEEPQSRKKLRMEAS DIP protein P61244 original sequence version: 132 DIP protein P61244 original sequence version: 128 DIP protein P61244 original sequence version: 150 DIP protein P61244 original sequence version: 120 EB10F3137727A56F skp2_human S-phase kinase-associated protein 2 F-box protein Skp2 Cyclin-A/CDK2-associated protein p45 p45skp2 F-box/LRR-repeat protein 1 SKP2 FBXL1 protein protein human Homo sapiens Human MHRKHLQEIPDLSSNVATSFTWGWDSSKTSELLSGMGVSALEKEEPDSENIPQELLSNLGHPESPPRKRLKSKGSDKDFVIVRRPKLNRENFPGVSWDSLPDELLLGIFSCLCLPELLKVSGVCKRWYRLASDESLWQTLDLTGKNLHPDVTGRLLSQGVIAFRCPRSFMDQPLAEHFSPFRVQHMDLSNSVIEVSTLHGILSQCSKLQNLSLEGLRLSDPIVNTLAKNSNLVRLNLSGCSGFSEFALQTLLSSCSRLDELNLSWCFDFTEKHVQVAVAHVSETITQLNLSGYRKNLQKSDLSTLVRRCPNLVHLDLSDSVMLKNDCFQEFFQLNYLQHLSLSRCYDIIPETLLELGEIPTLKTLQVFGIVPDGTLQLLKEALPHLQINCSHFTTIARPTIGNKKNQEIWGIKCRLTLQKPSCL F29B7C338A7A37E9 myc_human Myc proto-oncogene protein BHLHE39 Transcription factor p64 MYC Proto-oncogene c-Myc Class E basic helix-loop-helix protein 39 protein protein human Homo sapiens Human MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAPSEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTELLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPNPARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDSLLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGSPSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNRKCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKATAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA DIP protein P01106 original sequence version: 195 DIP protein P01106 original sequence version: 196 DIP protein P01106 original sequence version: 148 DIP protein P01106-1 original sequence version: 161 DIP protein P01106 original sequence version: 182 DIP protein P01106 original sequence version: 197 DIP protein P01106-1 original sequence version: 208 DIP protein P01106 original sequence version: 121 DIP protein P01106-1 original sequence version: 144 DIP protein P01106 original sequence version: 144 ED5C028029A4C5D1 pyr1_human_gene PYR1_HUMAN_GENE/CAD gene gene human Homo sapiens Human fbxw7_human F-box/WD repeat-containing protein 7 FBXW7 FBW7 FBX30 SEL10 F-box and WD-40 domain-containing protein 7 F-box protein FBX30 Archipelago homolog SEL-10 hCdc4 protein protein human Homo sapiens Human MNQELLSVGSKRRRTGGSLRGNPSSSQVDEEQMNRVVEEEQQQQLRQQEEEHTARNGEVVGVEPRPGGQNDSQQGQLEENNNRFISVDEDSSGNQEEQEEDEEHAGEQDEEDEEEEEMDQESDDFDQSDDSSREDEHTHTNSVTNSSSIVDLPVHQLSSPFYTKTTKMKRKLDHGSEVRSFSLGKKPCKVSEYTSTTGLVPCSATPTTFGDLRAANGQGQQRRRITSVQPPTGLQEWLKMFQSWSGPEKLLALDELIDSCEPTQVKHMMQVIEPQFQRDFISLLPKELALYVLSFLEPKDLLQAAQTCRYWRILAEDNLLWREKCKEEGIDEPLHIKRRKVIKPGFIHSPWKSAYIRQHRIDTNWRRGELKSPKVLKGHDDHVITCLQFCGNRIVSGSDDNTLKVWSAVTGKCLRTLVGHTGGVWSSQMRDNIIISGSTDRTLKVWNAETGECIHTLYGHTSTVRCMHLHEKRVVSGSRDATLRVWDIETGQCLHVLMGHVAAVRCVQYDGRRVVSGAYDFMVKVWDPETETCLHTLQGHTNRVYSLQFDGIHVVSGSLDTSIRVWDVETGNCIHTLTGHQSLTSGMELKDNILVSGNADSTVKIWDIKTGQCLQTLQGPNKHQSAVTCLQFNKNFVITSSDDGTVKLWDLKTGEFIRNLVTLESGGSGGVVWRIRASNTKLVCAVGSRNGTEETKLLVLDFDVDMK DIP protein Q969H0 original sequence version: 61 DIP protein Q969H0 original sequence version: 70 DIP protein Q969H0 original sequence version: 94 DIP protein Q969H0-1 original sequence version: 85 DIP protein Q969H0 original sequence version: 104 DIP protein Q969H0 original sequence version: 130 DIP protein Q969H0-1 original sequence version: 135 E4A357F76DFD8203 ep300_human Histone acetyltransferase p300 Histone butyryltransferase p300 Histone crotonyltransferase p300 Protein propionyltransferase p300 EP300 P300 E1A-associated protein p300 Protein 2-hydroxyisobutyryltransferase p300 Protein lactyltransferas p300 protein protein human Homo sapiens Human MAENVVEPGPPSAKRPKLSSPALSASASDGTDFGSLFDLEHDLPDELINSTELGLTNGGDINQLQTSLGMVQDAASKHKQLSELLRSGSSPNLNMGVGGPGQVMASQAQQSSPGLGLINSMVKSPMTQAGLTSPNMGMGTSGPNQGPTQSTGMMNSPVNQPAMGMNTGMNAGMNPGMLAAGNGQGIMPNQVMNGSIGAGRGRQNMQYPNPGMGSAGNLLTEPLQQGSPQMGGQTGLRGPQPLKMGMMNNPNPYGSPYTQNPGQQIGASGLGLQIQTKTVLSNNLSPFAMDKKAVPGGGMPNMGQQPAPQVQQPGLVTPVAQGMGSGAHTADPEKRKLIQQQLVLLLHAHKCQRREQANGEVRQCNLPHCRTMKNVLNHMTHCQSGKSCQVAHCASSRQIISHWKNCTRHDCPVCLPLKNAGDKRNQQPILTGAPVGLGNPSSLGVGQQSAPNLSTVSQIDPSSIERAYAALGLPYQVNQMPTQPQVQAKNQQNQQPGQSPQGMRPMSNMSASPMGVNGGVGVQTPSLLSDSMLHSAINSQNPMMSENASVPSLGPMPTAAQPSTTGIRKQWHEDITQDLRNHLVHKLVQAIFPTPDPAALKDRRMENLVAYARKVEGDMYESANNRAEYYHLLAEKIYKIQKELEEKRRTRLQKQNMLPNAAGMVPVSMNPGPNMGQPQPGMTSNGPLPDPSMIRGSVPNQMMPRITPQSGLNQFGQMSMAQPPIVPRQTPPLQHHGQLAQPGALNPPMGYGPRMQQPSNQGQFLPQTQFPSQGMNVTNIPLAPSSGQAPVSQAQMSSSSCPVNSPIMPPGSQGSHIHCPQLPQPALHQNSPSPVPSRTPTPHHTPPSIGAQQPPATTIPAPVPTPPAMPPGPQSQALHPPPRQTPTPPTTQLPQQVQPSLPAAPSADQPQQQPRSQQSTAASVPTPTAPLLPPQPATPLSQPAVSIEGQVSNPPSTSSTEVNSQAIAEKQPSQEVKMEAKMEVDQPEPADTQPEDISESKVEDCKMESTETEERSTELKTEIKEEEDQPSTSATQSSPAPGQSKKKIFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDIVKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQEIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQGESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAGFVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHASDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPPPNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDYIFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFEGDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLSRGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDLMDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQDRFVYTCNECKHHVETRWHCTVCEDYDLCITCYNTKNHDHKMEKLGLGLDDESNNQQAAATQSPGDSRRLSIQRCIQSLVHACQCRNANCSLPSCQKMKRVVQHTKGCKRKTNGGCPICKQLIALCCYHAKHCQENKCPVPFCLNIKQKLRQQQLQHRLQQAQMLRRRMASMQRTGVVGQQQGLPSPTPATPTTPTGQQPTTPQTPQPTSQPQPTPPNSMPPYLPRTQAAGPVSQGKAAGQVTPPTPPQTAQPPLPGPPPAAVEMAMQIQRAAETQRQMAHVQIFQRPIQHQMPPMTPMAPMGMNPPPMTRGPSGHLEPGMGPTGMQQQPPWSQGGLPQPQQLQSGMPRPAMMSVAQHGQPLNMAPQPGLGQVGISPLKPGTVSQQALQNLLRTLRSPSSPLQQQQVLSILHANPQLLAAFIKQRAAKYANSNPQPIPGQPGMPQGQPGLQPPTMPGQQGVHSNPAMQNMNPMQAGVQRAGLPQQQPQQQLQPPMGGMSPQAQQMNMNHNTMPSQFRDILRRQQMMQQQQQQGAGPGIGPGMANHNQFQQPQGVGYPPQQQQRMQHHMQQMQQGNMGQIGQLPQALGAEAGASLQAYQQRLLQQQMGSPVQPNPMSPQQHMLPNQAQSPHLQGQQIPNSLSNQVRSPQPVPSPRPQSQPPHSSPSPRMQPQPSPHHVSPQTSSPHPGLVAAQANPMEQGHFASPDQNSMLSQLASNPGMANLHGASATDLGLSTDNSDLNSNLSQSTLDIH DIP protein Q09472 original sequence version: 193 DIP protein Q09472 original sequence version: 186 DIP protein Q09472 original sequence version: 107 DIP protein Q09472 original sequence version: 112 DIP protein Q09472 original sequence version: 181 DIP protein Q09472 original sequence version: 183 DIP protein Q09472 original sequence version: 136 DIP protein Q09472 original sequence version: 146 DIP protein Q09472 original sequence version: 165 DIP protein Q09472 original sequence version: 192 DIP protein Q09472 original sequence version: 200 DIP protein Q09472 original sequence version: 206 DIP protein Q09472 original sequence version: 194 DIP protein Q09472 original sequence version: 171 DIP protein Q09472 original sequence version: 142 DIP protein Q09472 original sequence version: 180 8E869E1F174A6FEB ccnb1_human_gene G2/mitotic-specific cyclin-B1 gene gene gene human Homo sapiens Human snip1-myc 6 8 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey physical association physical association We performed a yeast two-hybrid screen with full-length SNIP1 and a HeLa cDNA library to identify SNIP1 partners. Nine of 21 strong positive clones encoded c-Myc, which did not interact with the N terminus of SNIP1 also used as bait. myc-snip1 5 8 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 11 unspecified role unspecified role bait bait physical association physical association 1 A, 5 C, 6 A We confirmed the interaction between c-Myc and SNIP1 in HEK293 cells using transiently overexpressed c-Myc and HA-tagged SNIP1 P01106; Q8TAD8 transfection P01106; Q8TAD8 overexpressed Binding of SNIP1 to c-Myc was abolished with Skp2 overexpression. The interaction of HA-SNIP1 with both endogenous c-Myc and ectopic c-Myc is enhanced by coexpression of Flag-p300. When the cells are not tranfected with a c-Myc expression vector HA-SNIP1 can interact with endogenous c-Myc. snip1-myc-1 2 11 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 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 1 A, 2D We confirmed the interaction between c-Myc and SNIP1 in HEK293 cells using transiently overexpressed c-Myc and HA-tagged SNIP1 P01106; Q8TAD8 transfection P01106; Q8TAD8 overexpressed snip1-myc-2 7 8 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey physical association physical association 1 B We confirmed the interaction between c-Myc and SNIP1 in HEK293 cells using transiently overexpressed c-Myc and HA-tagged SNIP1 (Figure 1A) as well as endogenous proteins myc-snip1-1 7 11 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association 1 B We confirmed the interaction between c-Myc and SNIP1 in HEK293 cells using transiently overexpressed c-Myc and HA-tagged SNIP1 (Figure 1A) as well as endogenous proteins max-snip1 7 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association 1 C The Max protein, a major component of the c-Myc transactivation complex (Bouchard et al., 1998), also bound SNIP1, max-myc-4 7 11 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association 1 C The Max protein, a major component of the c-Myc transactivation complex snip1-myc-3 4 11 unspecified role unspecified role prey prey in vitro In vitro 8 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 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association 2 A, B the fusion protein SNIP1 (215-321), which contains part of the FHA domain, bound to in vitro-translated c-Myc with the greatest affinity. The SNIP1 (215-321) construct also contains amino acids N terminal to the FHA domain. To test whether the FHA domain alone was sufficient to mediate binding, we used the GST fusion protein SNIP1 (274-349) containing only the FHA domain, and again showed binding to c-Myc, although this interaction is weaker compared to the 215-321 construct. myc-snip1-2 4 11 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 in vitro In vitro 8 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association 2 C Neither SNIP1-N (1-121) nor a SNIP1 construct that lacks the FHA domain-bound c-Myc. snip1-myc-4 7 8 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain physical association physical association 2 G HEK293 cell transfection experiments using c-Myc deletion mutants showed that D43 and DS c-Myc mutants interacted with endogenous SNIP1, whereas D147 c-Myc again failed to interact P01106 transfected myc-nt_022184_14 3 12 unspecified role unspecified role prey prey promoter_region CAD promoter E box domain sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role bait bait association association 3 G, 6D This suggests recruitment of SNIP1 and p300 to the Cad promoter is c-Myc dependent and that recruitment of p300 and SNIP1 is interdependent. It fits with cooperative regulation of c-Myc-dependent transcription by SNIP1 and p300. c-Myc siRNA greatly reduced this interaction. snip1-nt_022184_14-1 3 8 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey promoter_region CAD promoter E box domain sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association 3 G, 6 D Two antibodies raised against different domains of SNIP1 and a c-Myc antibody each immunoprecipitated the cyclin B1 and Cad promoter E box domains (Figure 3F) in control cells, but in c-Myc siRNA cells, promoter binding by both c-Myc and SNIP1 was greatly reduced. c-Myc siRNA or p300 siRNA greatly reduced this interaction. This suggests recruitment of SNIP1 and p300 to the Cad promoter is c-Myc dependent and that recruitment of p300 and SNIP1 is interdependent. It fits with cooperative regulation of c-Myc-dependent transcription by SNIP1 and p300. snip1-nt_006713_14 3 8 unspecified role unspecified role bait bait 15 unspecified role unspecified role prey prey promoter_region cyclin B promoter E box domain sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association 3 G Two antibodies raised against different domains of SNIP1 and a c-Myc antibody each immunoprecipitated the cyclin B1 and Cad promoter E box domains (Figure 3F) in control cells, but in c-Myc siRNA cells, promoter binding by both c-Myc and SNIP1 was greatly reduced. c-Myc siRNA greatly reduced this interaction. myc-nt_006713_14-1 3 11 unspecified role unspecified role bait bait 15 unspecified role unspecified role prey prey promoter_region cyclin B promoter E box domain sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association 3 G Two antibodies raised against different domains of SNIP1 and a c-Myc antibody each immunoprecipitated the cyclin B1 and Cad promoter E box domains (Figure 3F) in control cells, but in c-Myc siRNA cells, promoter binding by both c-Myc and SNIP1 was greatly reduced. c-Myc siRNA greatly reduced this interaction. myc-skp2 5 11 unspecified role unspecified role bait bait 10 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 5 B Overexpression of SNIP1 reduced Skp2 bound to c-Myc, concomitant with formation of a SNIP1/c-Myc complex P01106; Q13309 transfection P01106; Q13309 overexpressed Overexpression of SNIP1 reduced Skp2 bound to c-Myc. myc-fbxw7 5 11 unspecified role unspecified role bait bait 13 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 5 D P01106; Q969H0 transfection P01106; Q969H0 overexpressed Binding of Fbw7 to c-Myc was not affected by SNIP1 overexpression. binding of Fbw7 to c-Myc was unaffected by overexpressing SNIP1 myc-snip1-3 1 8 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association 5 F Shortly after serum addition, the interaction between c-Myc and SNIP1 was observed, but later in the cell cycle, as Skp2 levels began to rise, the c-Myc/ Skp2 complex became predominant This suggests Skp2 and SNIP1 compete for binding to c-Myc. myc-skp2-1 1 11 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association 5 F Shortly after serum addition, the interaction between c-Myc and SNIP1 was observed, but later in the cell cycle, as Skp2 levels began to rise, the c-Myc/ Skp2 complex became predominant This suggests Skp2 and SNIP1 compete for binding to c-Myc. myc-ep300 5 11 unspecified role unspecified role bait bait 14 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 6 A P01106; Q09472 transfection P01106; Q09472 overexpressed The interaction of Flag-p300 with both endogenous c-Myc and ectopic c-Myc is enhanced by coexpression of HA-SNIP1. When the cells are not tranfected with a c-Myc expression vector Flag-p300 can interact with endogenous c-Myc. the amounts of both the exogenous (lane 4 versus lane 8, short exposure) and endogenous (lane 2 versus lane 6, longer exposure) c-Myc/p300 complexes were enhanced by ectopic expression of SNIP1. ep300-nt_022184_14 3 12 unspecified role unspecified role prey prey promoter_region CAD promoter E box domain sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 14 unspecified role unspecified role bait bait association association 6 D c-Myc siRNA or SNIP1 siRNA greatly reduced this interaction. This suggests recruitment of SNIP1 and p300 to the Cad promoter is c-Myc dependent and that recruitment of p300 and SNIP1 is interdependent. It fits with cooperative regulation of c-Myc-dependent transcription by SNIP1 and p300. This experiment confirmed that recruitment of both SNIP1 and p300 to the Cad promoter is c-Myc dependent (Figure 6D). Significantly, recruitment of p300 and SNIP1 was found to be interdependent: depletion of either protein resulted in reduced recruitment of the other, without a significant reduction in c-Myc promoter binding.