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 Drosophila circadian network is a seasonal timer. The Drosophila circadian network is a seasonal timer. Cell (0092-8674) 2007 imex curation Stoleru D., Nawathean P., Fernandez Mde L., Menet JS., Ceriani MF., Rosbash M. rosbash@brandeis.edu 23-MAY-2007: Contacted by JYOTI. orchard Only protein-protein interactions 2 drome-head Drosophila melanogaster head anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Stoleru D., Nawathean P., Fernandez Mde L., Menet JS., Ceriani MF., Rosbash M. rosbash@brandeis.edu Accepted 2007-JUN-8 by ORCHARD. 23-MAY-2007: Contacted by JYOTI. Cell (0092-8674) 2007 orchard Only protein-protein interactions 2 imex curation The Drosophila circadian network is a seasonal timer. The Drosophila circadian network is a seasonal timer. Cell (0092-8674) 2007 imex curation Stoleru D., Nawathean P., Fernandez Mde L., Menet JS., Ceriani MF., Rosbash M. rosbash@brandeis.edu 23-MAY-2007: Contacted by JYOTI. orchard Only protein-protein interactions 2 drome-schneider_2 Drosophila melanogaster late embryo cells Fruit fly schneider_2 Established from the late embryo of Drosophila melanogaster Mixture of diploid and tetraploid. Originally had some XY cells now predominantly XX. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Stoleru D., Nawathean P., Fernandez Mde L., Menet JS., Ceriani MF., Rosbash M. rosbash@brandeis.edu Accepted 2007-JUN-8 by ORCHARD. 23-MAY-2007: Contacted by JYOTI. Cell (0092-8674) 2007 orchard Only protein-protein interactions 2 imex curation cry1_drome Cryptochrome-1 cry Blue light photoreceptor CG3772 protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MATRGANVIWFRHGLRLHDNPALLAALADKDQGIALIPVFIFDGESAGTKNVGYNRMRFLLDSLQDIDDQLQAATDGRGRLLVFEGEPAYIFRRLHEQVRLHRICIEQDCEPIWNERDESIRSLCRELNIDFVEKVSHTLWDPQLVIETNGGIPPLTYQMFLHTVQIIGLPPRPTADARLEDATFVELDPEFCRSLKLFEQLPTPEHFNVYGDNMGFLAKINWRGGETQALLLLDERLKVEQHAFERGFYLPNQALPNIHDSPKSMSAHLRFGCLSVRRFYWSVHDLFKNVQLRACVRGVQMTGGAHITGQLIWREYFYTMSVNNPNYDRMEGNDICLSIPWAKPNENLLQSWRLGQTGFPLIDGAMRQLLAEGWLHHTLRNTVATFLTRGGLWQSWEHGLQHFLKYLLDADWSVCAGNWMWVSSSAFERLLDSSLVTCPVALAKRLDPDGTYIKQYVPELMNVPKEFVHEPWRMSAEQQEQYECLIGVHYPERIIDLSMAVKRNMLAMKSLRNSLITPPPHCRPSNEEEVRQFFWLADVVV DIP protein O77059 original sequence version: 45 AB0019E5447BF56E sgg_drome Protein kinase shaggy sgg zw3 CG2621 Protein zeste-white 3 gsk3 protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MSGRPRTSSFAEGNKQSPSLVLGGVKTCSRDGSKITTVVATPGQGTDRVQEVSYTDTKVIGNGSFGVVFQAKLCDTGELVAIKKVLQDRRFKNRELQIMRKLEHCNIVKLLYFFYSSGEKRDEVFLNLVLEYIPETVYKVARQYAKTKQTIPINFIRLYMYQLFRSLAYIHSLGICHRDIKPQNLLLDPETAVLKLCDFGSAKQLLHGEPNVSYICSRYYRAPELIFGAINYTTKIDVWSAGCVLAELLLGQPIFPGDSGVDQLVEVIKVLGTPTREQIREMNPNYTEFKFPQIKSHPWQKVFRIRTPTEAINLVSLLLEYTPSARITPLKACAHPFFDELRMEGNHTLPNGRDMPPLFNFTEHELSIQPSLVPQLLPKHLQNASGPGGNRPSAGGAASIAASGSTSVSSTGSGASVEGSAQPQSQGTAAAAGSGSGGATAGTGGASAGGPGSGNNSSSGGASGAPSAVAAGGANAAVAGGAGGGGGAGAATAAATATGAIGATNAGGANVTDS DIP protein P18431 original sequence version: 169 193EE2A1294BE494 sgg-cry 2 4 unspecified role unspecified role bait bait region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 3 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 2 D We then determined that the CRY-SGG interaction was direct both in transgenic flies and in S2 cells. First, SGGV5, CRY-His6, or both were overexpressed in S2 cells, immunoprecipitated with anti-V5 antibody, Nickel beads, or both, and probed for CRY, SGG, or both, respectively O77059; P18431 tranfection O77059; P18431 over-expression cry-sgg 1 4 unspecified role unspecified role prey prey 3 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 physical association physical association 2 E Immunoprecipitation of CRY from fly head extracts with anti-MYC antibody followed by western blot with anti-SGG antibody validated the CRY-SGG interaction