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/ fenton-2004-1 Identification of the E1A-regulated transcription factor p120 E4F as an interacting partner of the RASSF1A candidate tumor suppressor gene. 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 Fenton SL., Dallol A., Agathanggelou A., Hesson L., Ahmed-Choudhury J., Baksh S., Sardet C., Dammann R., Minna JD., Downward J., Maher ER., Latif F. Apoptosis - Interactions involving proteins with a function related to apoptosis Note the UniProtKB entry Q66K89 for E4F1 has a higher degree of similarity to NP_004415 (the identifier given by the authors) than the other unreviewed entry of 784 aa. flatif@hgmp.mrc.ac.uk A MATCHMAKER human adult brain cDNA library (BD-Clontech) was used. Cancer Res. (0008-5472) 2004 Cancer - Interactions investigated in the context of cancer fenton-2004-2 Identification of the E1A-regulated transcription factor p120 E4F as an interacting partner of the RASSF1A candidate tumor suppressor gene. in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Fenton SL., Dallol A., Agathanggelou A., Hesson L., Ahmed-Choudhury J., Baksh S., Sardet C., Dammann R., Minna JD., Downward J., Maher ER., Latif F. Apoptosis - Interactions involving proteins with a function related to apoptosis Note the UniProtKB entry Q66K89 for E4F1 has a higher degree of similarity to NP_004415 (the identifier given by the authors) than the other unreviewed entry of 784 aa. flatif@hgmp.mrc.ac.uk A MATCHMAKER human adult brain cDNA library (BD-Clontech) was used. Cancer Res. (0008-5472) 2004 Cancer - Interactions investigated in the context of cancer fenton-2004-3 Identification of the E1A-regulated transcription factor p120 E4F as an interacting partner of the RASSF1A candidate tumor suppressor gene. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Fenton SL., Dallol A., Agathanggelou A., Hesson L., Ahmed-Choudhury J., Baksh S., Sardet C., Dammann R., Minna JD., Downward J., Maher ER., Latif F. Apoptosis - Interactions involving proteins with a function related to apoptosis Note the UniProtKB entry Q66K89 for E4F1 has a higher degree of similarity to NP_004415 (the identifier given by the authors) than the other unreviewed entry of 784 aa. flatif@hgmp.mrc.ac.uk A MATCHMAKER human adult brain cDNA library (BD-Clontech) was used. Cancer Res. (0008-5472) 2004 Cancer - Interactions investigated in the context of cancer fenton-2004-4 Identification of the E1A-regulated transcription factor p120 E4F as an interacting partner of the RASSF1A candidate tumor suppressor gene. mouse-3t3 Mus musculus embryo cells 3t3 Mouse embryo fibroblast cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Fenton SL., Dallol A., Agathanggelou A., Hesson L., Ahmed-Choudhury J., Baksh S., Sardet C., Dammann R., Minna JD., Downward J., Maher ER., Latif F. Apoptosis - Interactions involving proteins with a function related to apoptosis Note the UniProtKB entry Q66K89 for E4F1 has a higher degree of similarity to NP_004415 (the identifier given by the authors) than the other unreviewed entry of 784 aa. flatif@hgmp.mrc.ac.uk A MATCHMAKER human adult brain cDNA library (BD-Clontech) was used. Cancer Res. (0008-5472) 2004 Cancer - Interactions investigated in the context of cancer fenton-2004-5 Identification of the E1A-regulated transcription factor p120 E4F as an interacting partner of the RASSF1A candidate tumor suppressor gene. human-a_549 Homo sapiens Caucasian lung carcinoma cell line a_549 Human Caucasian lung carcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Fenton SL., Dallol A., Agathanggelou A., Hesson L., Ahmed-Choudhury J., Baksh S., Sardet C., Dammann R., Minna JD., Downward J., Maher ER., Latif F. Apoptosis - Interactions involving proteins with a function related to apoptosis Note the UniProtKB entry Q66K89 for E4F1 has a higher degree of similarity to NP_004415 (the identifier given by the authors) than the other unreviewed entry of 784 aa. flatif@hgmp.mrc.ac.uk A MATCHMAKER human adult brain cDNA library (BD-Clontech) was used. Cancer Res. (0008-5472) 2004 Cancer - Interactions investigated in the context of cancer fenton-2004-6 Identification of the E1A-regulated transcription factor p120 E4F as an interacting partner of the RASSF1A candidate tumor suppressor gene. human-nci_h1792 Homo sapiens non-small cell lung adenocarcinoma cell line NCI-H1792 nci_h1792 human non-small cell lung adenocarcinoma cell line ATCC number is CRL-5895. Tumour stage 4. Derived from pleural effusion. ATCC number is CRL-5895. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Fenton SL., Dallol A., Agathanggelou A., Hesson L., Ahmed-Choudhury J., Baksh S., Sardet C., Dammann R., Minna JD., Downward J., Maher ER., Latif F. Apoptosis - Interactions involving proteins with a function related to apoptosis Note the UniProtKB entry Q66K89 for E4F1 has a higher degree of similarity to NP_004415 (the identifier given by the authors) than the other unreviewed entry of 784 aa. flatif@hgmp.mrc.ac.uk A MATCHMAKER human adult brain cDNA library (BD-Clontech) was used. Cancer Res. (0008-5472) 2004 Cancer - Interactions investigated in the context of cancer e4f1_human Transcription factor E4F1 E4F1 E4F Putative E3 ubiquitin-protein ligase E4F1 E4F transcription factor 1 Transcription factor E4F p120E4F p50E4F RING-type E3 ubiquitin transferase E4F1 protein protein human Homo sapiens Human MEGAMAVRVTAAHTAEAQAEAGREAGEGAVAAVAAALAPSGFLGLPAPFSEEDEDDVHRCGRCQAEFTALEDFVQHKIQKACQRAPPEALPATPATTALLGQEVVPAAPGPEEPITVAHIVVEAASLAADISHASDLVGGGHIKEVIVAAEAELGDGEMAEAPGSPRQQGLGLAGEGEQAQVKLLVNKDGRYVCALCHKTFKTGSILKAHMVTHSSRKDHECKLCGASFRTKGSLIRHHRRHTDERPYKCSKCGKSFRESGALTRHLKSLTPCTEKIRFSVSKDVVVSKEDARAGSGAGAAGLGTATSSVTGEPIETSPVIHLVTDAKGTVIHEVHVQMQELSLGMKALAPEPPVSQELPCSSEGSRENLLHQAMQNSGIVLERAAGEEGALEPAPAAGSSPQPLAVAAPQLPVLEVQPLETQVASEASAVPRTHPCPQCSETFPTAATLEAHKRGHTGPRPFACAQCGKAFPKAYLLKKHQEVHVRERRFRCGDCGKLYKTIAHVRGHRRVHSDERPYPCPKCGKRYKTKNAQQVHFRTHLEEKPHVCQFCSRGFREKGSLVRHVRHHTGEKPFKCYKCGRGFAEHGTLNRHLRTKGGCLLEVEELLVSEDSPAAATTVLTEDPHTVLVEFSSVVADTQEYIIEATADDAETSEATEIIEGTQTEVDSHIMKVVQQIVHQASAGHQIIVQNVTMDEETALGPEAAAADTITIATPESLTEQVAMTLASAISEGTVLAARAGTSGTEQATVTMVSSEDIEILEHAGELVIASPEGQLEVQTVIV q9ns23-2 Ras association domain-containing protein 1 RASSF1A RASSF1 RDA32 protein protein human Homo sapiens Human MSGEPELIELRELAPAGRAGKGRTRLERANALRIARGTACNPTRQLVPGRGHRFQPAGPATHTWCDLCGDFIWGVVRKGLQCAHCKFTCHYRCRALVCLDCCGPRDLGWEPAVERDTNVDEPVEWETPDLSQAEIEQKIKEYNAQINSNLFMSLNKDGSYTGFIKVQLKLVRPVSVPSSKKPPSLQDARRGPGRGTSVRRRTSFYLPKDAVKHLHVLSRTRAREVIEALLRKFLVVDDPRKFALFERAERHGQVYLRKLLDDEQPLRLRLLAGPSDKALSFVLKENDSGEVNWDAFSMPELHNFLRILQREEEEHLRQILQKYSYCRQKIQEALHACPLG Produced by alternative splicing of isoform D. stk4_human Serine/threonine-protein kinase 4 STK4 MST1 STE20-like kinase MST1 Mammalian STE20-like protein kinase 1 Serine/threonine-protein kinase Krs-2 KRS2 protein protein human Homo sapiens Human METVQLRNPPRRQLKKLDEDSLTKQPEEVFDVLEKLGEGSYGSVYKAIHKETGQIVAIKQVPVESDLQEIIKEISIMQQCDSPHVVKYYGSYFKNTDLWIVMEYCGAGSVSDIIRLRNKTLTEDEIATILQSTLKGLEYLHFMRKIHRDIKAGNILLNTEGHAKLADFGVAGQLTDTMAKRNTVIGTPFWMAPEVIQEIGYNCVADIWSLGITAIEMAEGKPPYADIHPMRAIFMIPTNPPPTFRKPELWSDNFTDFVKQCLVKSPEQRATATQLLQHPFVRSAKGVSILRDLINEAMDVKLKRQESQQREVDQDDEENSEEDEMDSGTMVRAVGDEMGTVRVASTMTDGANTMIEHDDTLPSQLGTMVINAEDEEEEGTMKRRDETMQPAKPSFLEYFEQKEKENQINSFGKSVPGPLKNSSDWKIPQDGDYEFLKSWTVEDLQKRLLALDPMMEQEIEEIRQKYQSKRQPILDAIEAKKRRQQNF stk3_human Serine/threonine-protein kinase 3 STE20-like kinase MST2 Mammalian STE20-like protein kinase 2 Serine/threonine-protein kinase Krs-1 STK3 MST2 KRS1 protein protein human Homo sapiens Human MEQPPAPKSKLKKLSEDSLTKQPEEVFDVLEKLGEGSYGSVFKAIHKESGQVVAIKQVPVESDLQEIIKEISIMQQCDSPYVVKYYGSYFKNTDLWIVMEYCGAGSVSDIIRLRNKTLIEDEIATILKSTLKGLEYLHFMRKIHRDIKAGNILLNTEGHAKLADFGVAGQLTDTMAKRNTVIGTPFWMAPEVIQEIGYNCVADIWSLGITSIEMAEGKPPYADIHPMRAIFMIPTNPPPTFRKPELWSDDFTDFVKKCLVKNPEQRATATQLLQHPFIKNAKPVSILRDLITEAMEIKAKRHEEQQRELEEEEENSDEDELDSHTMVKTSVESVGTMRATSTMSEGAQTMIEHNSTMLESDLGTMVINSEDEEEEDGTMKRNATSPQVQRPSFMDYFDKQDFKNKSHENCNQNMHEPFPMSKNVFPDNWKVPQDGDFDFLKNLSLEELQMRLKALDPMMEREIEELRQRYTAKRQPILDAMDAKKRRQQNF rassf1-e4f1 RASSF1A interacts with E4F in a yeast two-hybrid assay 600060 N/A 600066 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind range range range range false N/A 600067 unspecified role unspecified role bait bait n-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false PREY protein NP_004415 Isoform C of RASS1F as described by the authors lacks 119 aa at the N-terminus as compared to RASSF1A and did not interact with E4F1 (461-784). Note that the isoform C of RASS1F described in the UniProtKB entry Q9NS23 does not conform with the isoform described by the authors. The authors state E4F is 683 aa in length, whereas the protein they reference (NP_004415) is 784 aa, as is the corresponding UniProtKB entry. The predicted C-terminal zinc finger range for NP_004415 is aa 416-595. The range given above for E4FDelta is approximate only as precise mapping was not possible as the sequence of E4FDelta was not provided. rassf1-stk4 RASSF1A interacts with MST1 in a yeast two-hybrid assay 600060 N/A 600067 unspecified role unspecified role bait bait N/A 600068 unspecified role unspecified role prey prey physical association physical association false false false rassf1-stk3 RASSF1A interacts with MST2 in a yeast two-hybrid assay 600060 N/A 600067 unspecified role unspecified role bait bait N/A 600069 unspecified role unspecified role prey prey physical association physical association false false false rassf1-e4f1-1 RASSF1A co-immunoprecipitates E4F in vitro 600061 N/A 600066 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind range range range range false region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 600067 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 false in vitro In vitro direct interaction direct interaction false false false PREY protein NP_004415 1 B e4f1-rassf1 RASSF1A co-immunoprecipitates with E4F in vitro 600061 N/A 600067 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 600066 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind range range range range false region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro direct interaction direct interaction false false false BAIT protein NP_004415 1 B rassf1-e4f1-2 RASSF1A pulls E4F in vitro 600062 N/A 600066 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region required to bind necessary binding region required to bind range range range range false in vitro In vitro N/A 600067 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false PREY protein NP_004415 1 C, 4 A The range for E4F is approximate only. It is based on the end of the fourth C-terminal zinc finger (found using zinc finger consensus sequence CX2-4C....HX2-4H) and the next 26 aa (as the deletion analysis reveals the 26 aa after the end of this fourth zinc finger are required to bind.) rassf1-e4f1-3 RASSF1A coimmunoprecipitates E4F in NIH 3T3 cells 600063 N/A 600067 unspecified role unspecified role bait bait N/A 600066 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false 2 A Q66K89; Q9NS23 transfection Q66K89; Q9NS23 over-expressed rassf1-e4f1-4 RASSF1A coimmunoprecipitates E4F in A549 cells 600064 N/A 600066 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 600067 unspecified role unspecified role bait bait physical association physical association false false false 2 A Q66K89; Q9NS23 transfection Q66K89; Q9NS23 over-expressed rassf1a-e4f1 Endogenous RASSF1A coimmunoprecipitates E4F1 in NCI-H1792 cells 600065 N/A 600066 unspecified role unspecified role prey prey N/A 600067 unspecified role unspecified role bait bait physical association physical association false false false 2 B, C This interaction is inhibited by RASSF1A-siRNA.