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/ http://www.ebi.ac.uk/ schmidt-2008-1 Characterization of the interaction between the human DNA topoisomerase IIbeta-binding protein 1 (TopBP1) and the cell division cycle 45 (Cdc45) protein. human-hela_s3 Homo sapiens HeLa-S3 cervix carcinoma cell line hela_s3 HeLa-S3 cervix carcinoma cell line Human cervix carcinoma cell line is a subclone of its parent HELA (DSM ACC 57) derived in 1955 confirmed as human with IEF of AST, MDH anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Schmidt U., Wollmann Y., Franke C., Grosse F., Saluz HP., Hanel F. Cancer - Interactions investigated in the context of cancer frank.haenel@hki-jena.de The stage of the cell cycle Biochem. J. (0264-6021) 2008 Only protein-protein interactions imex curation schmidt-2008-2 Characterization of the interaction between the human DNA topoisomerase IIbeta-binding protein 1 (TopBP1) and the cell division cycle 45 (Cdc45) protein. in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Schmidt U., Wollmann Y., Franke C., Grosse F., Saluz HP., Hanel F. Cancer - Interactions investigated in the context of cancer frank.haenel@hki-jena.de Biochem. J. (0264-6021) 2008 Only protein-protein interactions imex curation cdc45_human Cell division control protein 45 homolog CDC45 CDC45L2 UNQ374/PRO710 PORC-PI-1 CDC45L protein protein human Homo sapiens Human MFVSDFRKEFYEVVQSQRVLLFVASDVDALCACKILQALFQCDHVQYTLVPVSGWQELETAFLEHKEQFHYFILINCGANVDLLDILQPDEDTIFFVCDTHRPVNVVNVYNDTQIKLLIKQDDDLEVPAYEDIFRDEEEDEEHSGNDSDGSEPSEKRTRLEEEIVEQTMRRRQRREWEARRRDILFDYEQYEYHGTSSAMVMFELAWMLSKDLNDMLWWAIVGLTDQWVQDKITQMKYVTDVGVLQRHVSRHNHRNEDEENTLSVDCTRISFEYDLRLVLYQHWSLHDSLCNTSYTAARFKLWSVHGQKRLQEFLADMGLPLKQVKQKFQAMDISLKENLREMIEESANKFGMKDMRVQTFSIHFGFKHKFLASDVVFATMSLMESPEKDGSGTDHFIQALDSLSRSNLDKLYHGLELAKKQLRATQQTIASCLCTNLVISQGPFLYCSLMEGTPDVMLFSRPASLSLLSKHLLKSFVCSTKNRRCKLLPLVMAAPLSMEHGTVTVVGIPPETDSSDRKNFFGRAFEKAAESTSSRMLHNHFDLSVIELKAEDRSKFLDALISLLS topb1_human DNA topoisomerase 2-binding protein 1 KIAA0259 TOPBP1 DNA topoisomerase II-binding protein 1 DNA topoisomerase II-beta-binding protein 1 protein protein human Homo sapiens Human MSRNDKEPFFVKFLKSSDNSKCFFKALESIKEFQSEEYLQIITEEEALKIKENDRSLYICDPFSGVVFDHLKKLGCRIVGPQVVIFCMHHQRCVPRAEHPVYNMVMSDVTISCTSLEKEKREEVHKYVQMMGGRVYRDLNVSVTHLIAGEVGSKKYLVAANLKKPILLPSWIKTLWEKSQEKKITRYTDINMEDFKCPIFLGCIICVTGLCGLDRKEVQQLTVKHGGQYMGQLKMNECTHLIVQEPKGQKYECAKRWNVHCVTTQWFFDSIEKGFCQDESIYKTEPRPEAKTMPNSSTPTSQINTIDSRTLSDVSNISNINASCVSESICNSLNSKLEPTLENLENLDVSAFQAPEDLLDGCRIYLCGFSGRKLDKLRRLINSGGGVRFNQLNEDVTHVIVGDYDDELKQFWNKSAHRPHVVGAKWLLECFSKGYMLSEEPYIHANYQPVEIPVSHKPESKAALLKKKNSSFSKKDFAPSEKHEQADEDLLSQYENGSSTVVEAKTSEARPFNDSTHAEPLNDSTHISLQEENQSSVSHCVPDVSTITEEGLFSQKSFLVLGFSNENESNIANIIKENAGKIMSLLSRTVADYAVVPLLGCEVEATVGEVVTNTWLVTCIDYQTLFDPKSNPLFTPVPVMTGMTPLEDCVISFSQCAGAEKESLTFLANLLGASVQEYFVRKSNAKKGMFASTHLILKERGGSKYEAAKKWNLPAVTIAWLLETARTGKRADESHFLIENSTKEERSLETEITNGINLNSDTAEHPGTRLQTHRKTVVTPLDMNRFQSKAFRAVVSQHARQVAASPAVGQPLQKEPSLHLDTPSKFLSKDKLFKPSFDVKDALAALETPGRPSQQKRKPSTPLSEVIVKNLQLALANSSRNAVALSASPQLKEAQSEKEEAPKPLHKVVVCVSKKLSKKQSELNGIAASLGADYRWSFDETVTHFIYQGRPNDTNREYKSVKERGVHIVSEHWLLDCAQECKHLPESLYPHTYNPKMSLDISAVQDGRLCNSRLLSAVSSTKDDEPDPLILEENDVDNMATNNKESAPSNGSGKNDSKGVLTQTLEMRENFQKQLQEIMSATSIVKPQGQRTSLSRSGCNSASSTPDSTRSARSGRSRVLEALRQSRQTVPDVNTEPSQNEQIIWDDPTAREERARLASNLQWPSCPTQYSELQVDIQNLEDSPFQKPLHDSEIAKQAVCDPGNIRVTEAPKHPISEELETPIKDSHLIPTPQAPSIAFPLANPPVAPHPREKIITIEETHEELKKQYIFQLSSLNPQERIDYCHLIEKLGGLVIEKQCFDPTCTHIVVGHPLRNEKYLASVAAGKWVLHRSYLEACRTAGHFVQEEDYEWGSSSILDVLTGINVQQRRLALAAMRWRKKIQQRQESGIVEGAFSGWKVILHVDQSREAGFKRLLQSGGAKVLPGHSVPLFKEATHLFSDLNKLKPDDSGVNIAEAAAQNVYCLRTEYIADYLMQESPPHVENYCLPEAISFIQNNKELGTGLSQKRKAPTEKNKIKRPRVH cdc45l-topbp1 Cdc45 coimmunoprecipitates TopBP1 in HeLa-S3 lysates in a cell cycle-dependent manner 693208 N/A 693210 unspecified role unspecified role bait bait N/A 693211 unspecified role unspecified role prey prey physical association physical association false false false Fig. 1 A This interaction varies in strength with cell cycle phase. TopBP1 could be co-precipitated with Cdc45 at the G1/S boundary (0 h) and 1 h after release from the mimosine-block. At later time points (3 and 6 h) when more than half of the cell population was in the S-phase co-precipitation, efficiency dramatically declined. Only weak co-precipitation could be obtained with nocodazole-treated cells at the G2/M boundary. G1/S boundary (0h and 1 h after release from the mimosine-block). topbp1-cdc45l Cdc45 coimmunoprecipitates with TopBP1 in HeLa-S3 lysates in a cell-cycle dependent manner 693208 N/A 693211 unspecified role unspecified role prey prey N/A 693210 unspecified role unspecified role bait bait physical association physical association false false false Fig. 1 E This interaction occurred in lysates from synchronized HeLa-S3 cells in early S-phase 3 h after release from the double TdR-block. Early S phase (3 h after release from the double TdR-block). cdc45l-topbp1-2 Cdc45 coimmunoprecipitates TopBP1 in HeLa-S3 lysates in a cell cycle-dependent manner 693208 N/A 693210 unspecified role unspecified role bait bait N/A 693211 unspecified role unspecified role prey prey physical association physical association false false false Fig. 1 A This interaction varies in strength with cell cycle phase. TopBP1 could be co-precipitated with Cdc45 at the G1/S boundary (0 h) and 1 h after release from the mimosine-block. At later time points (3 and 6 h) when more than half of the cell population was in the S-phase co-precipitation, efficiency dramatically declined. Only weak co-precipitation could be obtained with nocodazole-treated cells at the G2/M boundary. S phase (3 h and 6 h after release from the mimosine-block). cdc45l-topbp1-3 Cdc45 coimmunoprecipitates TopBP1 in HeLa-S3 lysates in a cell cycle-dependent manner 693208 N/A 693210 unspecified role unspecified role bait bait N/A 693211 unspecified role unspecified role prey prey physical association physical association false false false Fig. 1 C This interaction varies in strength with cell cycle phase. TopBP1 could be co-precipitated with Cdc45 at the G1/S boundary (0 h) and 1 h after release from the mimosine-block. At later time points (3 and 6 h) when more than half of the cell population was in the S-phase co-precipitation, efficiency dramatically declined. Only weak co-precipitation could be obtained with nocodazole-treated cells at the G2/M boundary. G2/M boundary (nocodazole-treated cells). cdc45l-topbp1-1 GST-Cdc45 pulls down TopBP1 in vitro 693209 N/A 693211 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 false in vitro In vitro N/A 693210 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 false n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Fig. 2 topbp1-cdc45l-1 GST-TopBP1 pulls down Cdc45 in vitro 693209 N/A 693211 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 false n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) N/A 693210 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 false in vitro In vitro direct interaction direct interaction false false false Fig. 3