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 GAGA factor of Drosophila interacts with SAP18, a Sin3-associated polypeptide. The GAGA factor of Drosophila interacts with SAP18, a Sin3-associated polypeptide. EMBO Rep. (1469-221X) 2000 imex curation Espinas ML., Canudas S., Fanti L., Pimpinelli S., Casanova J., Azorin F. Chromatin - Epigenetic interactions resulting in chromatin modulation fambmc@cid.csic.es 09-MAR-2007: Contacted by JYOTI. Drosophila 0 to 12 h embryonic cDNA library. Only protein-protein interactions in vitro In vitro pull down pull down predetermined predetermined participant predetermined Espinas ML., Canudas S., Fanti L., Pimpinelli S., Casanova J., Azorin F. Chromatin - Epigenetic interactions resulting in chromatin modulation fambmc@cid.csic.es Accepted 2007-MAR-27 by DAVET. 09-MAR-2007: Contacted by JYOTI. EMBO Rep. (1469-221X) 2000 Only protein-protein interactions imex curation The GAGA factor of Drosophila interacts with SAP18, a Sin3-associated polypeptide. The GAGA factor of Drosophila interacts with SAP18, a Sin3-associated polypeptide. EMBO Rep. (1469-221X) 2000 imex curation Espinas ML., Canudas S., Fanti L., Pimpinelli S., Casanova J., Azorin F. Chromatin - Epigenetic interactions resulting in chromatin modulation fambmc@cid.csic.es 09-MAR-2007: Contacted by JYOTI. Drosophila 0 to 12 h embryonic cDNA library. 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 Espinas ML., Canudas S., Fanti L., Pimpinelli S., Casanova J., Azorin F. Chromatin - Epigenetic interactions resulting in chromatin modulation fambmc@cid.csic.es Accepted 2007-MAR-27 by DAVET. 09-MAR-2007: Contacted by JYOTI. EMBO Rep. (1469-221X) 2000 Only protein-protein interactions imex curation The GAGA factor of Drosophila interacts with SAP18, a Sin3-associated polypeptide. The GAGA factor of Drosophila interacts with SAP18, a Sin3-associated polypeptide. EMBO Rep. (1469-221X) 2000 imex curation Espinas ML., Canudas S., Fanti L., Pimpinelli S., Casanova J., Azorin F. Chromatin - Epigenetic interactions resulting in chromatin modulation fambmc@cid.csic.es 09-MAR-2007: Contacted by JYOTI. Drosophila 0 to 12 h embryonic cDNA library. 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 Espinas ML., Canudas S., Fanti L., Pimpinelli S., Casanova J., Azorin F. Chromatin - Epigenetic interactions resulting in chromatin modulation fambmc@cid.csic.es Accepted 2007-MAR-27 by DAVET. 09-MAR-2007: Contacted by JYOTI. Drosophila 0 to 12 h embryonic cDNA library. EMBO Rep. (1469-221X) 2000 Only protein-protein interactions imex curation gaga_drome Transcription factor GAGA Trl GAGA TFGAGA Trithorax-like protein GAGA factor Adh transcription factor 2 Neural conserved at 70F Adf-2 Nc70F CG33261 protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MSLPMNSLYSLTWGDYGTSLVSAIQLLRCHGDLVDCTLAAGGRSFPAHKIVLCAASPFLLDLLKNTPCKHPVVMLAGVNANDLEALLEFVYRGEVSVDHAQLPSLLQAAQCLNIQGLAPQTVTKDDYTTHSIQLQHMIPQHHDQDQLIATIATAPQQTVHAQVVEDIHHQGQILQATTQTNAAGQQQTIVTTDAAKHDQAVIQAFLPARKRKPRVKKMSPTAPKISKVEGMDTIMGTPTSSHGSGSVQQVLGENGAEGQLLSSTPIIKSEGQKVETIVTMDPNNMIPVTSANAATGEITPAQGATGSSGGNTSGVLSTPKAKRAKHPPGTEKPRSRSQSEQPATCPICYAVIRQSRNLRRHLELRHFAKPGVKKEKKTTSGKKSSSGSSGSGSGALSSSGSVPQVQTVQSLHTLQGVQVKKDPDAQQQQQQQQQQQQQQQQAMTVSGATGGQVQQQVQQVQQQVQQQQQQQQQQQQQLQHHQIIDSSGNITTATTSAQAAAAAQQQAAGQQQQLVAQSDGSESGAPLSIAQVQTLQGHQIIGNLNQVNMTDFQQQQPQQQQQQQQQQQQQQQQQQQTQQTL 04826A1B630E3320 sap18_drome Histone deacetylase complex subunit SAP18 Bin1 SAP18 CG6046 18 kDa Sin3-associated polypeptide Bicoid-interacting protein 1 dSAP18 protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MANVESMIVEEKTQVKQIDREKTCPMLLRVFCSTGRHHSVSEYMFGNVPTNELQIYTWQDATLHELTSLVRDVNPDTRKKGTYFDFAVVYPNFRSNHFQMREIGVTCTGQKGIDDNKTLAQAKFSIGDFLDISITPPNRLPPTARRQRPY 94947F206D8FA0AD q08605-1 Transcription factor GAGA F GAGA-581 Trl Adf-2 GAGA Nc70F TFGAGA Adh transcription factor 2 GAGA factor Neural conserved at 70F Trithorax-like protein CG33261 protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MSLPMNSLYSLTWGDYGTSLVSAIQLLRCHGDLVDCTLAAGGRSFPAHKIVLCAASPFLLDLLKNTPCKHPVVMLAGVNANDLEALLEFVYRGEVSVDHAQLPSLLQAAQCLNIQGLAPQTVTKDDYTTHSIQLQHMIPQHHDQDQLIATIATAPQQTVHAQVVEDIHHQGQILQATTQTNAAGQQQTIVTTDAAKHDQAVIQAFLPARKRKPRVKKMSPTAPKISKVEGMDTIMGTPTSSHGSGSVQQVLGENGAEGQLLSSTPIIKSEGQKVETIVTMDPNNMIPVTSANAATGEITPAQGATGSSGGNTSGVLSTPKAKRAKHPPGTEKPRSRSQSEQPATCPICYAVIRQSRNLRRHLELRHFAKPGVKKEKKTTSGKKSSSGSSGSGSGALSSSGSVPQVQTVQSLHTLQGVQVKKDPDAQQQQQQQQQQQQQQQQAMTVSGATGGQVQQQVQQVQQQVQQQQQQQQQQQQQLQHHQIIDSSGNITTATTSAQAAAAAQQQAAGQQQQLVAQSDGSESGAPLSIAQVQTLQGHQIIGNLNQVNMTDFQQQQPQQQQQQQQQQQQQQQQQQQTQQTL 04826A1B630E3320 q08605-2 Transcription factor GAGA C D E H GAGA-519a GAGA-519b Trl Adf-2 GAGA Nc70F TFGAGA Adh transcription factor 2 GAGA factor Neural conserved at 70F Trithorax-like protein CG33261 protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MSLPMNSLYSLTWGDYGTSLVSAIQLLRCHGDLVDCTLAAGGRSFPAHKIVLCAASPFLLDLLKNTPCKHPVVMLAGVNANDLEALLEFVYRGEVSVDHAQLPSLLQAAQCLNIQGLAPQTVTKDDYTTHSIQLQHMIPQHHDQDQLIATIATAPQQTVHAQVVEDIHHQGQILQATTQTNAAGQQQTIVTTDAAKHDQAVIQAFLPARKRKPRVKKMSPTAPKISKVEGMDTIMGTPTSSHGSGSVQQVLGENGAEGQLLSSTPIIKSEGQKVETIVTMDPNNMIPVTSANAATGEITPAQGATGSSGGNTSGVLSTPKAKRAKHPPGTEKPRSRSQSEQPATCPICYAVIRQSRNLRRHLELRHFAKPGVKKEKKSKSGNDTTLDSSMEMNTTAEGDNTVGSDGAGGAGSAGGQSSGTTPTRVISNAPQAAGAPAILAQGVLPQQQQQQQLQQQHQQHLTATLAGGGQAYIKHEGGGGGGTGQQQQQQAAQQQGMQNVIHIVGDQVFIPQQQQPQPQ Ref.2 (AAB81113/AAB81117) sequences are in conflict in position: 437:A->G. Ref.2 (AAB81113/AAB81117) sequences are in conflict in position: 437:A->G. 46EE082ACE1A0663 gaga-sap18 3 5 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait poz domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association For this screen, the first 245 amino acids of GAGA, which include the POZ domain (Figure 2A), were used. One of the 20 different clones isolated in this screen showed high homology to SAP18 gaga-sap18-1 2 7 unspecified role unspecified role bait bait poz domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role prey prey physical association physical association 1B However, the dSAP18, POZ245 interaction is weaker than the very strong homomeric POZ245, POZ245 interaction gaga-gaga 2 7 unspecified role unspecified role bait bait poz 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 poz domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 1b However, the dSAP18, POZ245 interaction is weaker than the very strong homomeric POZ245, POZ245 interaction gaga-sap18-2 1 7 unspecified role unspecified role prey prey 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) 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) physical association physical association 1C, 2B A GST SAP18 fusion protein was shown to interact very efficiently with recombinant GAGA The region of AA 122-519 did show some interaction but the region AA 122-280 did not interact with the SAP18. gaga-sap18-3 1 7 unspecified role unspecified role prey prey drome-sl2 Drosophila melanogaster sl1 cells sl2 drosophila sl12 cells 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) physical association physical association 1d As seen in Figure 1D, only the GAGA519 species appears to bind to GST dSAP18 with some efficiency. gaga-gaga-1 1 6 unspecified role unspecified role prey prey drome-sl2 Drosophila melanogaster sl1 cells sl2 drosophila sl12 cells 7 unspecified role unspecified role prey prey drome Drosophila melanogaster (Fruit fly) Fruit fly 7 unspecified role unspecified role bait bait poz domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) association association 1d Furthermore, a GST�POZ245 fusion was found to bind both GAGA forms indistinctly and much more efficiently gaga-sap18-4 1 5 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role bait bait region required to bind necessary binding region required 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 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) physical association physical association 1C, 2B A GST SAP18 fusion protein was shown to interact very efficiently with recombinant GAGA The region of AA 122-519 did show some interaction but the region AA 122-280 did not interact with the SAP18.