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/ Chfr interacts and colocalizes with TCTP to the mitotic spindle. Chfr interacts and colocalizes with TCTP to the mitotic spindle. Oncogene (0950-9232) 2008 imex curation Burgess A., Labbe JC., Vigneron S., Bonneaud N., Strub JM., Van Dorsselaer A., Lorca T., Castro A. Cancer - Interactions investigated in the context of cancer The paper (PMID 6425302) by one of the authors confirmed that in this paper they used Xenopus laevis egg extracts. thierry.lorca@crbm.cnrs.fr,anna.castro@crbm.cnrs.fr 27-Aug-2008: Contacted by JYOTI. jyoti 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 Burgess A., Labbe JC., Vigneron S., Bonneaud N., Strub JM., Van Dorsselaer A., Lorca T., Castro A. Cancer - Interactions investigated in the context of cancer The paper (PMID 6425302) by one of the authors confirmed that in this paper they used Xenopus laevis egg extracts. thierry.lorca@crbm.cnrs.fr,anna.castro@crbm.cnrs.fr Accepted 2008-AUG-4 by JYOTI. 27-Aug-2008: Contacted by JYOTI. A cDNA library constructed with human mRNAs isolated from three complete 6- to 8-week-old human embryos was used. Oncogene (0950-9232) 2008 jyoti Only protein-protein interactions imex curation Chfr interacts and colocalizes with TCTP to the mitotic spindle. Chfr interacts and colocalizes with TCTP to the mitotic spindle. Oncogene (0950-9232) 2008 imex curation Burgess A., Labbe JC., Vigneron S., Bonneaud N., Strub JM., Van Dorsselaer A., Lorca T., Castro A. Cancer - Interactions investigated in the context of cancer The paper (PMID 6425302) by one of the authors confirmed that in this paper they used Xenopus laevis egg extracts. thierry.lorca@crbm.cnrs.fr,anna.castro@crbm.cnrs.fr 27-Aug-2008: Contacted by JYOTI. jyoti Only protein-protein interactions xenla-oocyte Xenopus laevis oocyte Oocyte anti bait coip anti bait coimmunoprecipitation anti bait coip ms participant Identification by mass spectrometry ms participant Burgess A., Labbe JC., Vigneron S., Bonneaud N., Strub JM., Van Dorsselaer A., Lorca T., Castro A. Cancer - Interactions investigated in the context of cancer The paper (PMID 6425302) by one of the authors confirmed that in this paper they used Xenopus laevis egg extracts. thierry.lorca@crbm.cnrs.fr,anna.castro@crbm.cnrs.fr Accepted 2008-AUG-4 by JYOTI. Oncogene (0950-9232) 2008 jyoti Only protein-protein interactions imex curation Chfr interacts and colocalizes with TCTP to the mitotic spindle. Chfr interacts and colocalizes with TCTP to the mitotic spindle. Oncogene (0950-9232) 2008 imex curation Burgess A., Labbe JC., Vigneron S., Bonneaud N., Strub JM., Van Dorsselaer A., Lorca T., Castro A. Cancer - Interactions investigated in the context of cancer The paper (PMID 6425302) by one of the authors confirmed that in this paper they used Xenopus laevis egg extracts. thierry.lorca@crbm.cnrs.fr,anna.castro@crbm.cnrs.fr 27-Aug-2008: Contacted by JYOTI. jyoti Only protein-protein interactions xenla-oocyte Xenopus laevis oocyte Oocyte molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Burgess A., Labbe JC., Vigneron S., Bonneaud N., Strub JM., Van Dorsselaer A., Lorca T., Castro A. Cancer - Interactions investigated in the context of cancer Gel filtration was carried out on interphase egg extracts. The paper (PMID 6425302) by one of the authors confirmed that in this paper they used Xenopus laevis egg extracts. thierry.lorca@crbm.cnrs.fr,anna.castro@crbm.cnrs.fr Accepted 2008-AUG-4 by JYOTI. 27-Aug-2008: Contacted by JYOTI. Oncogene (0950-9232) 2008 jyoti Only protein-protein interactions imex curation Chfr interacts and colocalizes with TCTP to the mitotic spindle. Chfr interacts and colocalizes with TCTP to the mitotic spindle. Oncogene (0950-9232) 2008 imex curation Burgess A., Labbe JC., Vigneron S., Bonneaud N., Strub JM., Van Dorsselaer A., Lorca T., Castro A. Cancer - Interactions investigated in the context of cancer The paper (PMID 6425302) by one of the authors confirmed that in this paper they used Xenopus laevis egg extracts. thierry.lorca@crbm.cnrs.fr,anna.castro@crbm.cnrs.fr 27-Aug-2008: Contacted by JYOTI. jyoti Only protein-protein interactions xenla-xl2 Xenopus laevis tadpole cell line XL2 xl2 Xenopus laevis tadpole cell line XL2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Burgess A., Labbe JC., Vigneron S., Bonneaud N., Strub JM., Van Dorsselaer A., Lorca T., Castro A. Cancer - Interactions investigated in the context of cancer The paper (PMID 6425302) by one of the authors confirmed that in this paper they used Xenopus laevis egg extracts. thierry.lorca@crbm.cnrs.fr,anna.castro@crbm.cnrs.fr Accepted 2008-AUG-4 by JYOTI. 27-Aug-2008: Contacted by JYOTI. Oncogene (0950-9232) 2008 jyoti Only protein-protein interactions imex curation Chfr interacts and colocalizes with TCTP to the mitotic spindle. Chfr interacts and colocalizes with TCTP to the mitotic spindle. Oncogene (0950-9232) 2008 imex curation Burgess A., Labbe JC., Vigneron S., Bonneaud N., Strub JM., Van Dorsselaer A., Lorca T., Castro A. Cancer - Interactions investigated in the context of cancer The paper (PMID 6425302) by one of the authors confirmed that in this paper they used Xenopus laevis egg extracts. thierry.lorca@crbm.cnrs.fr,anna.castro@crbm.cnrs.fr 27-Aug-2008: Contacted by JYOTI. jyoti Only protein-protein interactions xenla-oocyte Xenopus laevis oocyte Oocyte anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Burgess A., Labbe JC., Vigneron S., Bonneaud N., Strub JM., Van Dorsselaer A., Lorca T., Castro A. Cancer - Interactions investigated in the context of cancer The paper (PMID 6425302) by one of the authors confirmed that in this paper they used Xenopus laevis egg extracts. thierry.lorca@crbm.cnrs.fr,anna.castro@crbm.cnrs.fr Accepted 2008-AUG-4 by JYOTI. 27-Aug-2008: Contacted by JYOTI. Oncogene (0950-9232) 2008 jyoti Only protein-protein interactions imex curation Chfr interacts and colocalizes with TCTP to the mitotic spindle. Chfr interacts and colocalizes with TCTP to the mitotic spindle. Oncogene (0950-9232) 2008 imex curation Burgess A., Labbe JC., Vigneron S., Bonneaud N., Strub JM., Van Dorsselaer A., Lorca T., Castro A. Cancer - Interactions investigated in the context of cancer The paper (PMID 6425302) by one of the authors confirmed that in this paper they used Xenopus laevis egg extracts. thierry.lorca@crbm.cnrs.fr,anna.castro@crbm.cnrs.fr 27-Aug-2008: Contacted by JYOTI. jyoti 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 Burgess A., Labbe JC., Vigneron S., Bonneaud N., Strub JM., Van Dorsselaer A., Lorca T., Castro A. Cancer - Interactions investigated in the context of cancer The paper (PMID 6425302) by one of the authors confirmed that in this paper they used Xenopus laevis egg extracts. thierry.lorca@crbm.cnrs.fr,anna.castro@crbm.cnrs.fr Accepted 2008-AUG-4 by JYOTI. 27-Aug-2008: Contacted by JYOTI. Oncogene (0950-9232) 2008 jyoti Only protein-protein interactions imex curation chfr_xenla E3 ubiquitin-protein ligase CHFR chfr Checkpoint with forkhead and RING finger domains protein RING-type E3 ubiquitin transferase CHFR protein protein xenla Xenopus laevis (African clawed frog) African clawed frog MEGLDEKKPWGKLSRLLGAETDSSSELFLYKKEWTIGRKKACDLSFPGNKLVSGEHCKITVNEESGEVSLEDTSTNGTVINKLKVIRKQTYPLKNGDVIYVVYRKNEPEQNVAYLYKSLNQGQDSLHDPADTSGSEEAETQTLSSQDDQLSYEEPQPSTSTSSLFSTPTTSAIPGVQLESAEKSGESLGGHSSTSDASPAIRASIPKSNLSTQEQGSLGPPKKRIRTEDHWTTNKNFVPASCPIGASDESKTPSMKPDKMEETLTCIICQELLHDCVSLQPCMHTFCAACYSGWMERSSLCPTCRCPVERICKNHILNNLVEAYLIQHPEKCRSEEDRCSMDARNKITQDMLQPKVRRSFSDEEGSSEDLLELSDVDSESSDISQPYTVCRQCPGFVRHSMQPPPYPPPSDTETSRTQGDAPSTSTNFPTATQEYVCPSHGSHVICTCCFQPMPDRRAEREHNSHVAPQQCTICLEPFCHMYWGCNRMGCFGCLAPFCELNLGDKCLDGVLNNNNYESDILKNYLASRGLTWKDMLNESLAAVQRGVFMLPDYRINGTTVLCYFCGLRNFRILTYQYRQNIPASELPVTVTSRPNCYWGRNCRTQVKAHHAMKFNHICEQTRFKN 72C0A497D48F6BDC tctp_xenla Translationally-controlled tumor protein homolog tpt1 protein protein xenla Xenopus laevis (African clawed frog) African clawed frog MIIYKDCITEDEMFSDIYKIIETPDGMCLEVEGKVIQREEGAIDDALIGGNASAEFQEDDVGGSSLTSGVDIVMNHKLQETGFTKDSYKNYIKDYVKQLKAHLEKTNPERVNTFMKGAQETVKKILGNFKNYQFYTGERMNPDGMVGLLDYREDGITPFMIFFKDGLIIEKC 1F34F2B7B1E9D168 tctp_human Translationally-controlled tumor protein TPT1 p23 Histamine-releasing factor Fortilin protein protein human Homo sapiens Human MIIYRDLISHDEMFSDIYKIREIADGLCLEVEGKMVSRTEGNIDDSLIGGNASAEGPEGEGTESTVITGVDIVMNHHLQETSFTKEAYKKYIKDYMKSIKGKLEEQRPERVKPFMTGAAEQIKHILANFKNYQFFIGENMNPDGMVALLDYREDGVTPYMIFFKDGLEMEKC DIP protein P13693 original sequence version: 147 BD31399B71CA62F9 ubiqp_xenla Polyubiquitin protein protein xenla Xenopus laevis (African clawed frog) African clawed frog QKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGG DIP protein P62972 original sequence version: 41 9C8F431544081131 chfr_human E3 ubiquitin-protein ligase CHFR Checkpoint with forkhead and RING finger domains protein RING finger protein 196 CHFR RNF196 RING-type E3 ubiquitin transferase CHFR protein protein human Homo sapiens Human MERPEEGKQSPPPQPWGRLLRLGAEEGEPHVLLRKREWTIGRRRGCDLSFPSNKLVSGDHCRIVVDEKSGQVTLEDTSTSGTVINKLKVVKKQTCPLQTGDVIYLVYRKNEPEHNVAYLYESLSEKQGMTQESFEANKENVFHGTKDTSGAGAGRGADPRVPPSSPATQVCFEEPQPSTSTSDLFPTASASSTEPSPAGRERSSSCGSGGGGISPKGSGPSVASDEVSSFASALPDRKTASFSSLEPQDQEDLEPVKKKMRGDGDLDLNGQLLVAQPRRNAQTVHEDVRAAAGKPDKMEETLTCIICQDLLHDCVSLQPCMHTFCAACYSGWMERSSLCPTCRCPVERICKNHILNNLVEAYLIQHPDKSRSEEDVQSMDARNKITQDMLQPKVRRSFSDEEGSSEDLLELSDVDSESSDISQPYVVCRQCPEYRRQAAQPPHCPAPEGEPGAPQALGDAPSTSVSLTTAVQDYVCPLQGSHALCTCCFQPMPDRRAEREQDPRVAPQQCAVCLQPFCHLYWGCTRTGCYGCLAPFCELNLGDKCLDGVLNNNSYESDILKNYLATRGLTWKNMLTESLVALQRGVFLLSDYRVTGDTVLCYCCGLRSFRELTYQYRQNIPASELPVAVTSRPDCYWGRNCRTQVKAHHAMKFNHICEQTRFKN 141A1E7FEFAE36A2 chfr-tctp 1 9 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association Chfr were used as baits to screen a cDNA library obtained from a 6 to 8-week-old human embryo. One of the positive clones isolated was the translationally controlled tumor protein (TCTP). chfr-ubiq 2 7 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association 1 B The results showed that the focused band corresponded exclusively to Chfr, whereas Chfr and ubiquitin were present in all of the sequenced smeared bands, indicating that endogenous Chfr is continuously ubiquitinated in CSF extracts. chfr-tctp-1 6 11 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association Mating assays subsequently confirmed an interaction between LexA-Chfr and TCTP. chfr-tctp-2 3 8 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component physical association physical association 5 B Gel filtration performed on interphase extracts showed that a small amount of endogenous TCTP coeluted with Chfr. chfr-tctp-3 5 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association 5 C TCTP binds to Chfr in Xenopus egg extracts chfr-tctp-4 4 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association 5 C A weak but consistent band corresponding to TCTP was obtained in the Chfr immunoprecipitate from CSF and XL2 extracts