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/ Interaction of Sp1 with the growth- and cell cycle-regulated transcription factor E2F. Interaction of Sp1 with the growth- and cell cycle-regulated transcription factor E2F. Mol. Cell. Biol. (0270-7306) 1996 imex curation Karlseder J., Rotheneder H., Wintersberger E. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Wi@Mol.UniVie.AC.AT 10-APR-2007: Contacted by JYOTI. orchard Only protein-protein interactions mouse-3t3 Mus musculus embryo cells 3t3 Mouse embryo fibroblast cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Karlseder J., Rotheneder H., Wintersberger E. Apoptosis - Interactions involving proteins with a function related to apoptosis Wi@Mol.UniVie.AC.AT Accepted 2007-APR-19 by ORCHARD. 10-APR-2007: Contacted by JYOTI. Mol. Cell. Biol. (0270-7306) 1996 orchard Only protein-protein interactions imex curation Interaction of Sp1 with the growth- and cell cycle-regulated transcription factor E2F. Interaction of Sp1 with the growth- and cell cycle-regulated transcription factor E2F. Mol. Cell. Biol. (0270-7306) 1996 imex curation Karlseder J., Rotheneder H., Wintersberger E. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Wi@Mol.UniVie.AC.AT 10-APR-2007: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Karlseder J., Rotheneder H., Wintersberger E. Apoptosis - Interactions involving proteins with a function related to apoptosis Wi@Mol.UniVie.AC.AT Accepted 2007-APR-19 by ORCHARD. 10-APR-2007: Contacted by JYOTI. Mol. Cell. Biol. (0270-7306) 1996 orchard Only protein-protein interactions imex curation Interaction of Sp1 with the growth- and cell cycle-regulated transcription factor E2F. Interaction of Sp1 with the growth- and cell cycle-regulated transcription factor E2F. Mol. Cell. Biol. (0270-7306) 1996 imex curation Karlseder J., Rotheneder H., Wintersberger E. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Wi@Mol.UniVie.AC.AT 10-APR-2007: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Karlseder J., Rotheneder H., Wintersberger E. Apoptosis - Interactions involving proteins with a function related to apoptosis Wi@Mol.UniVie.AC.AT Accepted 2007-APR-19 by ORCHARD. 10-APR-2007: Contacted by JYOTI. Mol. Cell. Biol. (0270-7306) 1996 orchard Only protein-protein interactions imex curation e2f2_human Transcription factor E2F2 E2F2 protein protein human Homo sapiens Human MLQGPRALASAAGQTPKVVPAMSPTELWPSGLSSPQLCPATATYYTPLYPQTAPPAAAPGTCLDATPHGPEGQVVRCLPAGRLPAKRKLDLEGIGRPVVPEFPTPKGKCIRVDGLPSPKTPKSPGEKTRYDTSLGLLTKKFIYLLSESEDGVLDLNWAAEVLDVQKRRIYDITNVLEGIQLIRKKAKNNIQWVGRGMFEDPTRPGKQQQLGQELKELMNTEQALDQLIQSCSLSFKHLTEDKANKRLAYVTYQDIRAVGNFKEQTVIAVKAPPQTRLEVPDRTEDNLQIYLKSTQGPIEVYLCPEEVQEPDSPSEEPLPSTSTLCPSPDSAQPSSSTDPSIMEPTASSVPAPAPTPQQAPPPPSLVPLEATDSLLELPHPLLQQTEDQFLSPTLACSSPLISFSPSLDQDDYLWGLEAGEGISDLFDSYDLGDLLIN DIP protein Q14209 original sequence version: 133 60541F4235507005 e2f3_human Transcription factor E2F3 E2F3 KIAA0075 protein protein human Homo sapiens Human MRKGIQPALEQYLVTAGGGEGAAVVAAAAAASMDKRALLASPGFAAAAAAAAAPGAYIQILTTNTSTTSCSSSLQSGAVAAGPLLPSAPGAEQTAGSLLYTTPHGPSSRAGLLQQPPALGRGGSGGGGGPPAKRRLELGESGHQYLSDGLKTPKGKGRAALRSPDSPKTPKSPSEKTRYDTSLGLLTKKFIQLLSQSPDGVLDLNKAAEVLKVQKRRIYDITNVLEGIHLIKKKSKNNVQWMGCSLSEDGGMLAQCQGLSKEVTELSQEEKKLDELIQSCTLDLKLLTEDSENQRLAYVTYQDIRKISGLKDQTVIVVKAPPETRLEVPDSIESLQIHLASTQGPIEVYLCPEETETHSPMKTNNQDHNGNIPKPASKDLASTNSGHSDCSVSMGNLSPLASPANLLQQTEDQIPSNLEGPFVNLLPPLLQEDYLLSLGEEEGISDLFDAYDLEKLPLVEDFMCS DIP protein O00716 original sequence version: 137 4641565842CA99EC e2f1_human Transcription factor E2F1 Retinoblastoma-associated protein 1 Retinoblastoma-binding protein 3 PBR3 E2F1 RBBP3 pRB-binding protein E2F-1 protein protein human Homo sapiens Human MALAGAPAGGPCAPALEALLGAGALRLLDSSQIVIISAAQDASAPPAPTGPAAPAAGPCDPDLLLFATPQAPRPTPSAPRPALGRPPVKRRLDLETDHQYLAESSGPARGRGRHPGKGVKSPGEKSRYETSLNLTTKRFLELLSHSADGVVDLNWAAEVLKVQKRRIYDITNVLEGIQLIAKKSKNHIQWLGSHTTVGVGGRLEGLTQDLRQLQESEQQLDHLMNICTTQLRLLSEDTDSQRLAYVTCQDLRSIADPAEQMVMVIKAPPETQLQAVDSSENFQISLKSKQGPIDVFLCPEETVGGISPGKTPSQEVTSEEENRATDSATIVSPPPSSPPSSLTTDPSQSLLSLEQEPLLSRMGSLRAPVDEDRLSPLVAADSLLEHVREDFSGLLPEEFISLSPPHEALDYHFGLEEGEGIRDLFDCDFGDLTPLDF DIP protein Q01094 original sequence version: 173 DIP protein Q01094 original sequence version: 168 DIP protein Q01094 original sequence version: 149 DIP protein Q01094 original sequence version: 130 DIP protein Q01094 original sequence version: 125 DIP protein Q01094 original sequence version: 153 DIP protein Q01094 original sequence version: 180 003B3F654F0C60DF e2f1_mouse Transcription factor E2F1 E2f1 protein protein mouse Mus musculus Mouse MAVAPAGGQHAPALEALLGAGALRLLDSSQIVIISTAPDVGAPQLPAAPPTGPRDSDVLLFATPQAPRPAPSAPRPALGRPPVKRRLDLETDHQYLAGSSGPFRGRGRHPGKGVKSPGEKSRYETSLNLTTKRFLELLSRSADGVVDLNWAAEVLKVQKRRIYDITNVLEGIQLIAKKSKNHIQWLGSHTMVGIGKRLEGLTQDLQQLQESEQQLDHLMHICTTQLQLLSEDSDTQRLAYVTCQDLRSIADPAEQMVIVIKAPPETQLQAVDSSETFQISLKSKQGPIDVFLCPEESADGISPGKTSCQETSSGEDRTADSGPAGPPPSPPSTSPALDPSQSLLGLEQEAVLPRMGHLRVPMEEDQLSPLVAADSLLEHVKEDFSGLLPGEFISLSPPHEALDYHFGLEEGEGIRDLFDCDFGDLTPLDF C5DF18AD3B4DFEFA sp1_mouse Transcription factor Sp1 Sp1 Specificity protein 1 protein protein mouse Mus musculus Mouse MSDQDHSMDEVTAVVKIEKDVGGNNGGSGNGGGAAFSQTRSSSTGSSSSSGGGGGQESQPSPLALLAATCSRIESPNENSNNSQGPSQSGGTGELDLTATQLSQGANGWQIISSSSGATPTSKEQSGNSTNGSNGSESSKNRTVSGGQYVVAATPNLQNQQVLTGLPGVMPNIQYQVIPQFQTVDGQQLQFAATGAQVQQDGSGQIQIIPGANQQIIPNRGSGGNIIAAMPNLLQQAVPLQGLANNVLSGQTQYVTNVPVALNGNITLLPVNSVSAATLTPSSQAGTISSSGSQESSSQPVTSGTAISSASLVSSQASSSSFFTNANSYSTTTTTSNMGIMNFTSSGSSGTSSQGQTPQRVGGLQGSDSLNIQQNQTSGGSLQGSQQKEGEQSQQTQQQQILIQPQLVQGGQALQALQAAPLSGQTFTTQAISQETLQNLQLQAVQNSGPIIIRTPTVGPNGQVSWQTLQLQNLQVQNPQAQTITLAPMQGVSLGQTSSSNTTLTPIASAASIPAGTVTVNAAQLSSMPGLQTINLSALGTSGIQVHQLPGLPLAIANTPGDHGTQLGLHGSGGDGIHDETAGGEGENSSDLQPQAGRRTRREACTCPYCKDSEGRASGDPGKKKQHICHIQGCGKVYGKTSHLRAHLRWHTGERPFMCNWSYCGKRFTRSDELQRHKRTHTGEKKFACPECPKRFMRSDHLSKHIKTHQNKKGGPGVALSVGTLPLDSGAGSEGTATPSALITTNMVAMEAICPEGIARLANSGINVMQVTELQSINISGNGF DIP protein O89090 original sequence version: 112 B6B989F0A252CEE5 sp1_human Transcription factor Sp1 SP1 TSFP1 protein protein human Homo sapiens Human MSDQDHSMDEMTAVVKIEKGVGGNNGGNGNGGGAFSQARSSSTGSSSSTGGGGQESQPSPLALLAATCSRIESPNENSNNSQGPSQSGGTGELDLTATQLSQGANGWQIISSSSGATPTSKEQSGSSTNGSNGSESSKNRTVSGGQYVVAAAPNLQNQQVLTGLPGVMPNIQYQVIPQFQTVDGQQLQFAATGAQVQQDGSGQIQIIPGANQQIITNRGSGGNIIAAMPNLLQQAVPLQGLANNVLSGQTQYVTNVPVALNGNITLLPVNSVSAATLTPSSQAVTISSSGSQESGSQPVTSGTTISSASLVSSQASSSSFFTNANSYSTTTTTSNMGIMNFTTSGSSGTNSQGQTPQRVSGLQGSDALNIQQNQTSGGSLQAGQQKEGEQNQQTQQQQILIQPQLVQGGQALQALQAAPLSGQTFTTQAISQETLQNLQLQAVPNSGPIIIRTPTVGPNGQVSWQTLQLQNLQVQNPQAQTITLAPMQGVSLGQTSSSNTTLTPIASAASIPAGTVTVNAAQLSSMPGLQTINLSALGTSGIQVHPIQGLPLAIANAPGDHGAQLGLHGAGGDGIHDDTAGGEEGENSPDAQPQAGRRTRREACTCPYCKDSEGRGSGDPGKKKQHICHIQGCGKVYGKTSHLRAHLRWHTGERPFMCTWSYCGKRFTRSDELQRHKRTHTGEKKFACPECPKRFMRSDHLSKHIKTHQNKKGGPGVALSVGTLPLDSGAGSEGSGTATPSALITTNMVAMEAICPEGIARLANSGINVMQVADLQSINISGNGF DIP protein P08047 original sequence version: 185 DIP protein P08047 original sequence version: 187 DIP protein P08047 original sequence version: 181 DIP protein P08047 original sequence version: 162 DIP protein P08047 original sequence version: 161 43893DBF6518B9EA e2f1-sp1 2 6 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 n-terminal required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 8 unspecified role unspecified role prey prey mouse-3t3 Mus musculus embryo cells 3t3 Mouse embryo fibroblast cells physical association physical association 2 A, B, D, E As can be seen in Fig. 2A, E2F1 and truncated versions thereof retained Sp1 provided that the N-terminal 122 amino acids e2f1-sp1-1 2 6 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 ecobb Escherichia coli (strain B / BL21) 9 unspecified role unspecified role prey prey direct interaction direct interaction 2 A Purified Sp1 could be used in this assay in place of the Sp1-containing cell extract and gave the same results, indicating that the binding of Sp1 to E2F1 is direct and not mediated by additional cellular proteins. sp1-e2f1 3 9 unspecified role unspecified role bait bait c terminus required to bind necessary binding region required to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1212396]range positions must always be strictly superior to 0 and inferior or equal to the sequence length. Actual positions : 788-788, sequence length 785 [EBI-1212396]622-788 6 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 2 C This experiment (Fig. 2C) revealed that the binding domain for E2F within Sp1 lies within the C-terminal quarter of the protein The length of the human Sp1 sequence used here is given as 788 aa, however the UniProtKb sequence is only 785 aa. e2f2-sp1 2 4 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 ecobb Escherichia coli (strain B / BL21) 8 unspecified role unspecified role prey prey mouse-3t3 Mus musculus embryo cells 3t3 Mouse embryo fibroblast cells physical association physical association 2 E it is evident that, as expected, E2F1, E2F2, and E2F3 bound Sp1 whereas E2F4 and E2F5 failed to do so. Sequence comparison of E2F1-5 reveals some similarity between E2F1, E2F2, and E2F3 within that part of the molecules required for binding to Sp1. E2F4 and E2F5 lack this region almost completely and do not interact with Sp1. e2f3-sp1 2 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 8 unspecified role unspecified role prey prey mouse-3t3 Mus musculus embryo cells 3t3 Mouse embryo fibroblast cells physical association physical association 2 E it is evident that, as expected, E2F1, E2F2, and E2F3 bound Sp1 whereas E2F4 and E2F5 failed to do so. Sequence comparison of E2F1-5 reveals some similarity between E2F1, E2F2, and E2F3 within that part of the molecules required for binding to Sp1. E2F4 and E2F5 lack this region almost completely and do not interact with Sp1. e2f1-sp1-2 1 7 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association 3 Sp1 was bound by antibody against E2F1, indicating that complexes of the two proteins exist in vivo.