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/ Treslin collaborates with TopBP1 in triggering the initiation of DNA replication. Treslin collaborates with TopBP1 in triggering the initiation of DNA replication. Cell (0092-8674) 2010 imex curation Kumagai A., Shevchenko A., Shevchenko A., Dunphy WG. dunphy@caltech.edu 29-Mar-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 4 in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Kumagai A., Shevchenko A., Shevchenko A., Dunphy WG. dunphy@caltech.edu Accepted 2010-APR-16 by JYOTI. 29-Mar-2010: Contacted by JYOTI. Cell (0092-8674) 2010 jyoti Only protein-protein interactions 4 imex curation Treslin collaborates with TopBP1 in triggering the initiation of DNA replication. Treslin collaborates with TopBP1 in triggering the initiation of DNA replication. Cell (0092-8674) 2010 imex curation Kumagai A., Shevchenko A., Shevchenko A., Dunphy WG. dunphy@caltech.edu 29-Mar-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 4 xenla-oocyte Xenopus laevis oocyte Oocyte anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Kumagai A., Shevchenko A., Shevchenko A., Dunphy WG. dunphy@caltech.edu Accepted 2010-APR-16 by JYOTI. 29-Mar-2010: Contacted by JYOTI. Cell (0092-8674) 2010 jyoti Only protein-protein interactions 4 imex curation Treslin collaborates with TopBP1 in triggering the initiation of DNA replication. Treslin collaborates with TopBP1 in triggering the initiation of DNA replication. Cell (0092-8674) 2010 imex curation Kumagai A., Shevchenko A., Shevchenko A., Dunphy WG. dunphy@caltech.edu 29-Mar-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 4 in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip sequence tag sequence tag identification sequence tag Kumagai A., Shevchenko A., Shevchenko A., Dunphy WG. dunphy@caltech.edu Accepted 2010-APR-16 by JYOTI. 29-Mar-2010: Contacted by JYOTI. Cell (0092-8674) 2010 jyoti Only protein-protein interactions 4 imex curation ticrr_xenla Treslin TopBP1-interacting checkpoint and replication regulator TopBP1-interacting, replication-stimulating protein ticrr protein protein xenla Xenopus laevis (African clawed frog) African clawed frog MAPSHNVVLLVDTAESSDKSRLRRVSLRLLNFLACRAGLGQVRWSYRFLNSSGGRCRPPRRSDLRELGPRGWEEFEDELEACWERARNCRPSSTQSSRAQLLQTALMETLADFQWDRPDITSPTKPTLLRSRRGRIVAADEPLKDDSPDNFINPHSRNSIFLLSSCPHSGTELGQFAATSGDFSTQKVMDKLLPKSLQKIVSSKRVRVYWLDTSDWTQFGSSSDHSGYWTMVELMHQVEGRILPSESILGSSCQKAKTLPSFSVPPINIPFESVLNYLIFSEPDYQLWFPRRDGILFLTGKDGTKQLDCAVSLEPVSMIQKLSTSLMTIELKGTMQNCNLPLAGLRVETWLLHNSDCVQLQKLTKELMLKELHMIATVTLEDDVLPRTGILSPLSETAAVLNVICSERTLGLDNLHVQGSVHETDKETFSDLPDIVMSVLNHVYSSEDNTLAPDFPVPEWIKQELSQSSRWTSSVTARWYPLSGVSGASCNLMESFRLINAASSNCDEHLKFDQELTNYLSEFYQKKSVDDAGLGVHRENQKKSGLPRTPVRQKMKTLPRALQMLNAARLNVKAQKADSTLPVPNEKNSQMKRRSSGKQDNKPKQLKPTEFQSEDGLISYIKENYEEAVSLVDHSTMTWARDTLTTIKSYLKSIGSEQIETEAIDKVKLLFKTSKVIRQNYRNNQDKEVKLKECQLQVFLRLEMFVQCPVIQMDSDELELIIEEITDILRIVSLTEDPLFLTKFLEVDVLTQYIASVPKILADIYFSLGTQIPEVLVLVLPSDGDDSIMHEEKSVKSQPSTSRVPSVAPIGAETDQLEDLRTRSAKKRRSTALARHRSVAESSQSFRQIEVPKRQPNKENVQSNAVVVLEKLKLPLPAQPQKDAEAKVRRNLFIQETRSPSKRCSKMPRSQSVSAVESLKRKRSKSHDGSKDHHKLLTKKVSETPVHKQTANRLLLRQIKGRPSESNSNISIVEESPEKEIRDIDLRRSPRIKQLSLTRRNSSSFYASQPKSRNLERVNSATQLQQSRERPGSCLISEVKTPKRLLFGEVLGMISPPTTKRSRRILDMVNPVYKTPGKTPRKTPSKNIPNFEDQSGNMLVKSPCTPYTPRTPSRTPKRLKTPSKGSTERKKAAKNLGKLFSPSKPEEKSPLKLWGRRSERLAQMTPGKDGSPYKQSVCQTLMEVKTPQKLQRLESKDFRTPSRTPTRSNNTTPAKQSMQISNTPRKSDLKHPQEHESRGPSGYILWTPQKRILASVPHTPILQTPQKPISASVPRTPVCRTPQKAILSSVPSTPVCQTPNKAILTSVSRTPVYQTPQKAILASVMSTPVYQTPQKPVLASVPTTPILKTPQRSALASVSHTPSPKKYIMKELTVAITRMRECTPEKVLGSNLSSSATPSSALKSFCSEKTTSVCQTPKKSSIALLKPCDSLEFSGAPERLMDSLCSNKDSTKAETACTVPSQISTQMQNVINAGECTDSLSQTSVSSPSIPFTDKSLSPDLKDALSDVTSSKAEGVTIVSEKLDSSSMDSQEATDSFINSSQTEESIDISEARVVSTEASELKMKVLITRKPSGSGVSYLPTTPKCLGNVCSTSTYGLRCTPDRRQREAAARLGTPEIPAKFSTPKSHCKMIPQSIYEVELEMQESGLPKLRFKRTDSNSTIDMDVNKTPKISRKRKGDESPFNEKWCSKHAVRTEPACVSPSCVRTSHYTPGKSGIQTFICQSYTPNRCLSAAASPSQSDAGVPWTPSPKEKLSTDVINSWPRKKKASALCTNLLKCDKIPEYAEEDGGDFELEGVSKLLEKSPVIEQQSKVDGGTFGLRSRKRVFSLVSPTKETENPVKRVCTFNRHEDSSTATHRHQTKEEMEIFSSDQSRSSYLSSSQQSICDDVFNMSDFTPPSKVPKNPLSACGLLTLTQSPLLYKGKTPSSKRKEKIQDVFSDGDSDHGTPTLKRPTNPAAVSDDSPFRKVNPLRSISKTYSRKKLIT FE3B4C817FD867C5 tob1a_xenla DNA topoisomerase 2-binding protein 1-A Cut5 protein DNA topoisomerase II-binding protein 1-A topbp1-A cut5 protein protein xenla Xenopus laevis (African clawed frog) African clawed frog MASSENEPFCVKFIKSPENSEYFFKAYEAIKQIQSDESLQLTEEREALLLKEKDKSLYICDPFSGAAFSHLKKLGCRIVGPQVVIFCMENQRRVPRAEYPVYNMAMADVTISCTSLDKETREDVHHYVQIMGGCVYRDLNVSVTHLIAGEVGSKKYLVAASLEKPILLPSWVKELWEKSNQRIIRYSDVNMTEYLCPIFRGCTICVTGLSSLDRKEVQRLTALHGGEYTGQLKMNESTHLIVQEAKGQKYECARKWNVHCISVQWFFDSIEKGFCQDETMYKIEPASTIKSVPDTSTPTGGNSKPNSRTLYDVSQISNISTSCVNESAFNSAMASRLDPPADTLENLDISSLQAPDDLLDGCRIYLCGFGGRKLDKLRKLINNGGGVRFNQLTGDVTHIIVGETDEELKQFLNKTQHRPYVLTVKWLLDSFAKGHLQPEEIYFHSSYQQTEMPSPFEPAINLTANKMSSTRDPLNHTRNHQADEDLLSQYTENNSTLIEDEHPKTSNTNSISQMSMHEDMTTCTSQSGLADTSTIIEGGLFSRKQFMVLGFLEEDEACIIDIIKKSAGKVLSSQKRAIADYAVVPLLGCEVESTVGEVVTNAWLGMCIEQEKLLDPHSNALFTPVPFLEGSTPLRECVLSVSQFMGAERDSLVYLAGLLGAKVQEFFVRKANPKKGMFASTHLVLKDAEGSKYEAAKKWNLPAVTMNWLLQCARTGRKADEDSYLVDNVPEEDKDESFISQTYKPQAIRLSMHAPCHLENHPEALTKAAVTPLDMNRFKSKAFQSVISQHNKNPQTSGGESKVLQREPSLHLDTPSKFLSKDKLFKPSFDVKDALAALETPGGPNQKNRTQSTPLSEVIGRNLQLAIANSTRQTAAVTASPQLKAAEKKEFDNSKLLINVVICVSKKLIKKQGELNGIAASLGAEYRWCFDESVTHFIYHGRQNDMSREYKSVKERSGIYIVSEHWLFACSEQQKRVPEALYPHTYNPKMSLDISAVQDGSYTASKFSADTSLQQDENSELQLQQNNKFGETSDDQVKKAAGDGNPQNPSKDVKGALTQTLEMRENFQRQLQEFMSATSVVKPRGSVGRAGFDNSPCTPEGARSTRNGRSRVLEALRQSRQAMTDLNTEPSQNEQIIWDDPTAREERAKLVSNLQWPDSPSQYSEQLQHNMNDAGGNYTPAKESLTDTEIAELEACEFEPKSAMRTPVIENNLQSPTKPDHLTPTPQAPSIAFPLANPPVAPQPREKPVQPFSKEETLKERRFQLSSLDPQERIDYSQLIEELGGVVIEKQCFDPSCTHIVVGHPLRNEKYLASMAAGKWVLHRSYLEACRAAKRFIQEEDYEWGSMSILSAVTNINPQQRMLAEAAMRWRKKLQGIKQNMGIAEGAFSGWKVILNVDQTKEPGFKRLLQSGGAKVFAGHSSPLFKEASHLFADFSKLKPDEPRVNVAEAAAQGVNCLKPEYIADYLMKELPPPMNNYCLPDAIPYVRVTGTGLSRKRKTSGDVSDVKRSRHY 31BB4CB37DEF225B topbp1-loc443616 3 4 unspecified role unspecified role prey prey xenla-oocyte Xenopus laevis oocyte Oocyte 5 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal 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 c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection physical association physical association Fig. 1A We have identified a protein called Treslin that associates with TopBP1 in Xenopus egg extracts. topbp1-loc443616-1 1 5 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal 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 c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 4 unspecified role unspecified role prey prey xenla-oocyte Xenopus laevis oocyte Oocyte physical association physical association Fig. 1C, 2D, 2E, 4C We could readily detect binding of endogenous Treslin to beads containing the tagged TopBP1 but not to control beads p27 CDKN1B topbp1-loc443616-2 2 5 unspecified role unspecified role bait bait spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 4 unspecified role unspecified role prey prey xenla-oocyte Xenopus laevis oocyte Oocyte physical association physical association Fig. 4A, Supp. Fig. 3 and anti-TopBP1 immunoprecipitates (lanes 4, 5, 7, and 8) were immunoblotted for Treslin and TopBP1. p27 CDKN1B Binding of Treslin to TopBP1 increases substantially near the commencement of DNA replication, triggered by addition of sperm nuclei extracts. topbp1-loc443616-3 2 5 unspecified role unspecified role prey prey spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 4 unspecified role unspecified role bait bait xenla-oocyte Xenopus laevis oocyte Oocyte physical association physical association Fig. 4A and anti-TopBP1 immunoprecipitates (lanes 4, 5, 7, and 8) were immunoblotted for Treslin and TopBP1. p27 CDKN1B