MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein. The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein. Oncogenesis 2019 imex curation Zarich N., Anta B., Fernández-Medarde A., Ballester A., de Lucas MP., Cámara AB., Anta B., Oliva JL., Rojas-Cabañeros JM., Santos E. Interactions in fig. 2b and 2c were not annotated since anti-CSN7a antibody is able to immunoprecipate the whole COP9 complex jloliva@isciii.es; jmrojas@isciii.es; esantos@usal.es 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zarich N., Anta B., Fernández-Medarde A., Ballester A., de Lucas MP., Cámara AB., Anta B., Oliva JL., Rojas-Cabañeros JM., Santos E. Interactions in fig. 2b and 2c were not annotated since anti-CSN7a antibody is able to immunoprecipate the whole COP9 complex jloliva@isciii.es; jmrojas@isciii.es; esantos@usal.es Accepted 2019-MAR-11 AT 13:31 GMT AT 13:31 GMT by LUANA Oncogenesis 2019 Only protein-protein interactions imex curation imex curation The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein. The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein. Oncogenesis 2019 imex curation Zarich N., Anta B., Fernández-Medarde A., Ballester A., de Lucas MP., Cámara AB., Anta B., Oliva JL., Rojas-Cabañeros JM., Santos E. Interactions in fig. 2b and 2c were not annotated since anti-CSN7a antibody is able to immunoprecipate the whole COP9 complex jloliva@isciii.es; jmrojas@isciii.es; esantos@usal.es 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Zarich N., Anta B., Fernández-Medarde A., Ballester A., de Lucas MP., Cámara AB., Anta B., Oliva JL., Rojas-Cabañeros JM., Santos E. Interactions in fig. 2b and 2c were not annotated since anti-CSN7a antibody is able to immunoprecipate the whole COP9 complex jloliva@isciii.es; jmrojas@isciii.es; esantos@usal.es Accepted 2019-MAR-11 AT 13:31 GMT AT 13:31 GMT by LUANA Oncogenesis 2019 Only protein-protein interactions imex curation imex curation The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein. The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein. Oncogenesis 2019 imex curation Zarich N., Anta B., Fernández-Medarde A., Ballester A., de Lucas MP., Cámara AB., Anta B., Oliva JL., Rojas-Cabañeros JM., Santos E. Interactions in fig. 2b and 2c were not annotated since anti-CSN7a antibody is able to immunoprecipate the whole COP9 complex jloliva@isciii.es; jmrojas@isciii.es; esantos@usal.es 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-embryonic_fibroblast Mus musculus embryonic fibroblasts embryonic_fibroblast Embryonic fibroblast Fibroblasts taken from an embryonic as opposed to a foetal or adult source. Foetus is defined as the stage at which tissue diferentiation is initiated. Tissue taken from an embryonic as opposed to a foetal or adult source. Embryo is defined as the development stage prior to organ formation. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zarich N., Anta B., Fernández-Medarde A., Ballester A., de Lucas MP., Cámara AB., Anta B., Oliva JL., Rojas-Cabañeros JM., Santos E. Interactions in fig. 2b and 2c were not annotated since anti-CSN7a antibody is able to immunoprecipate the whole COP9 complex jloliva@isciii.es; jmrojas@isciii.es; esantos@usal.es Accepted 2019-MAR-11 AT 13:31 GMT AT 13:31 GMT by LUANA Oncogenesis 2019 Only protein-protein interactions imex curation imex curation The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein. The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein. Oncogenesis 2019 imex curation Zarich N., Anta B., Fernández-Medarde A., Ballester A., de Lucas MP., Cámara AB., Anta B., Oliva JL., Rojas-Cabañeros JM., Santos E. Interactions in fig. 2b and 2c were not annotated since anti-CSN7a antibody is able to immunoprecipate the whole COP9 complex jloliva@isciii.es; jmrojas@isciii.es; esantos@usal.es 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Zarich N., Anta B., Fernández-Medarde A., Ballester A., de Lucas MP., Cámara AB., Anta B., Oliva JL., Rojas-Cabañeros JM., Santos E. Interactions in fig. 2b and 2c were not annotated since anti-CSN7a antibody is able to immunoprecipate the whole COP9 complex jloliva@isciii.es; jmrojas@isciii.es; esantos@usal.es Accepted 2019-MAR-11 AT 13:31 GMT AT 13:31 GMT by LUANA Oncogenesis 2019 Only protein-protein interactions imex curation imex curation csn3_mouse COP9 signalosome complex subunit 3 JAB1-containing signalosome subunit 3 Cops3 Csn3 protein protein mouse Mus musculus Mouse MASALEQFVNSVRQLSAQGQMTQLCELINKSGELLAKNLSHLDTVLGALDVQEHSLGVLAVLFVKFSMPSVPDFETLFSQVQLFISTCNGEHIRYATDTFAGLCHQLTNALVERKQPLRGIGILKQAIDKMQMNTNQLTSVHADLCQLCLLAKCFKPALPYLDVDMMDICKENGAYDAKHFLCYYYYGGMIYTGLKNFERALYFYEQAITTPAMAVSHIMLESYKKYILVSLILLGKVQQLPKYTSQIVGRFIKPLSNAYHELAQVYSTNNPSELRNLVSKHSETFTRDNNMGLVKQCLSSLYKKNIQRLTKTFLTLSLQDMASRVQLSGPQEAEKYVLHMIEDGEIFASINQKDGMVSFHDNPEKYNNPAMLHNIDQEMLKCIELDERLKAMDQEITVNPQFVQKSMGSQEDDSGNKPSSYS 2FDCCDB98A5168FB sos1_human Son of sevenless homolog 1 SOS1 protein protein human Homo sapiens Human MQAQQLPYEFFSEENAPKWRGLLVPALKKVQGQVHPTLESNDDALQYVEELILQLLNMLCQAQPRSASDVEERVQKSFPHPIDKWAIADAQSAIEKRKRRNPLSLPVEKIHPLLKEVLGYKIDHQVSVYIVAVLEYISADILKLVGNYVRNIRHYEITKQDIKVAMCADKVLMDMFHQDVEDINILSLTDEEPSTSGEQTYYDLVKAFMAEIRQYIRELNLIIKVFREPFVSNSKLFSANDVENIFSRIVDIHELSVKLLGHIEDTVEMTDEGSPHPLVGSCFEDLAEELAFDPYESYARDILRPGFHDRFLSQLSKPGAALYLQSIGEGFKEAVQYVLPRLLLAPVYHCLHYFELLKQLEEKSEDQEDKECLKQAITALLNVQSGMEKICSKSLAKRRLSESACRFYSQQMKGKQLAIKKMNEIQKNIDGWEGKDIGQCCNEFIMEGTLTRVGAKHERHIFLFDGLMICCKSNHGQPRLPGASNAEYRLKEKFFMRKVQINDKDDTNEYKHAFEIILKDENSVIFSAKSAEEKNNWMAALISLQYRSTLERMLDVTMLQEEKEEQMRLPSADVYRFAEPDSEENIIFEENMQPKAGIPIIKAGTVIKLIERLTYHMYADPNFVRTFLTTYRSFCKPQELLSLIIERFEIPEPEPTEADRIAIENGDQPLSAELKRFRKEYIQPVQLRVLNVCRHWVEHHFYDFERDAYLLQRMEEFIGTVRGKAMKKWVESITKIIQRKKIARDNGPGHNITFQSSPPTVEWHISRPGHIETFDLLTLHPIEIARQLTLLESDLYRAVQPSELVGSVWTKEDKEINSPNLLKMIRHTTNLTLWFEKCIVETENLEERVAVVSRIIEILQVFQELNNFNGVLEVVSAMNSSPVYRLDHTFEQIPSRQKKILEEAHELSEDHYKKYLAKLRSINPPCVPFFGIYLTNILKTEEGNPEVLKRHGKELINFSKRRKVAEITGEIQQYQNQPYCLRVESDIKRFFENLNPMGNSMEKEFTDYLFNKSLEIEPRNPKPLPRFPKKYSYPLKSPGVRPSNPRPGTMRHPTPLQQEPRKISYSRIPESETESTASAPNSPRTPLTPPPASGASSTTDVCSVFDSDHSSPFHSSNDTVFIQVTLPHGPRSASVSSISLTKGTDEVPVPPPVPPRRRPESAPAESSPSKIMSKHLDSPPAIPPRQPTSKAYSPRYSISDRTSISDPPESPPLLPPREPVRTPDVFSSSPLHLQPPPLGKKSDHGNAFFPNSPSPFTPPPPQTPSPHGTRRHLPSPPLTQEVDLHSIAGPPVPPRQSTSQHIPKLPPKTYKREHTHPSMHRDGPPLLENAHSS DIP protein Q07889 original sequence version: 117 DIP protein Q07889 original sequence version: 116 DIP protein Q07889 original sequence version: 124 DIP protein Q07889 original sequence version: 171 C6B99CCA11A8DE45 csn3_human COP9 signalosome complex subunit 3 COPS3 CSN3 JAB1-containing signalosome subunit 3 protein protein human Homo sapiens Human MASALEQFVNSVRQLSAQGQMTQLCELINKSGELLAKNLSHLDTVLGALDVQEHSLGVLAVLFVKFSMPSVPDFETLFSQVQLFISTCNGEHIRYATDTFAGLCHQLTNALVERKQPLRGIGILKQAIDKMQMNTNQLTSIHADLCQLCLLAKCFKPALPYLDVDMMDICKENGAYDAKHFLCYYYYGGMIYTGLKNFERALYFYEQAITTPAMAVSHIMLESYKKYILVSLILLGKVQQLPKYTSQIVGRFIKPLSNAYHELAQVYSTNNPSELRNLVNKHSETFTRDNNMGLVKQCLSSLYKKNIQRLTKTFLTLSLQDMASRVQLSGPQEAEKYVLHMIEDGEIFASINQKDGMVSFHDNPEKYNNPAMLHNIDQEMLKCIELDERLKAMDQEITVNPQFVQKSMGSQEDDSGNKPSSYS DIP protein Q9UNS2 original sequence version: 82 DIP protein Q9UNS2-1 original sequence version: 122 DIP protein Q9UNS2 original sequence version: 140 1D371050C7D7BF8D sos1_mouse Son of sevenless homolog 1 Sos1 protein protein mouse Mus musculus Mouse MQAQQLPYEFFSEENAPKWRGLLVPALKKVQGQVHPTLESNDDALQYVEELILQLLNMLCQAQPRSASDVEERVQKSFPHPIDKWAIADAQSAIEKRKRRNPLSLPAERIHHLLREVLGYKIDHQVSVYIVAVLEYISADILKLVGNYVRNIRHYEITKQDIKVAMCADKVLMDMFHQDVEDINILSLTDEEPSTSGEQTYYDLVKAFMAEIRQYIRELNLIIKVFREPFVSNSKLFSSNDVENIFSRIVDIHELSVKLLGHIEDTVEMTDEGSPHPLVGSCFEDLAEELAFDPYESYARDILRPGFHGHFLSQLSKPGAALYLQSIGEGFKEAVQYVLPRLLLAPVYHCLHYFELLKQLEEKSEDQEDKECMKQAITALLNVQSGMEKICSKSLAKRRLSESACRFYSQQMKGKQLAIKKMNEIQKNIDGWEGKDIGQCCNEFIMEGTLTRVGAKHERHIFLFDGLMICCKSNHGQPRLPGASSAEYRLKEKFFMRKVQINDKDDTSEYKHAFEIILKDGNSVIFSAKSAEEKNNWMAALISLQYRSTLERMLDVTVLQEEKEEQMRLPSAEVYRFAEPDSEENILFEENVQPKAGIPIIKAGTVLKLIERLTYHMYADPNFVRTFLTTYRSFCRPQELLSLLIERFEIPEPEPTEADRIAIENGDQPLSAELKRFRKEYIQPVQLRVLNVCRHWVEHHFYDFERDADLLQRMEEFIGTVRGKAMKKWVESITKIIQRKKIARDNGPGHNITFQSSPPTVEWHISRPGHIETFDLLTLHPIEIARQLTLLESDLYRAVQPSELVGSVWTKEDKEINSPNLLKMIRHTTNLTLWFEKCIVETENLEERVAVVSRIIEILQVFQELNNFNGVLEVVSAMNSSPVYRLDHTFEQIPSRQKKILEEAHELSEDHYKKYLAKLRSINPPCVPFFGIYLTNILKTEEGNPEVLRRHGKELINFSKRRRVAEITGEIQQYQNQPYCLRVEPDIKRFFENLNPMGNSMEKEFTDYLFNKSLEIEPRHPKPLPRFPKKYSYPLKSPGVRPSNPRPGTMRHPTPLQQEPRKISYSRIPESETESTASAPNSPRTPLTPPPASGTSSNTDVCSVFDSDHSASPFHSRSASVSSISLSKGTDEVPVPPPVPPRRRPESAPAESSPSKIMSKHLDSPPAIPPRQPTSKAYSPRYSISDRTSISDPPESPPLLPPREPVRTPDVFSSSPLHLQPPPLGKKSDHGNAFFPNSPSPFTPPPPQTPSPHGTRRHLPSPPLTQEMDLHSIAGPPVPPRQSTSQLIPKLPPKTYKREHTHPSMHRDGPPLLENAHSS 3286088A5BA0A4A6 3286088A5BA0A4A6 sos1-cops3-1 2 6 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role prey prey cdna library cdna library physical association physical association fig.1 Initial screening using a LexA-NDP plasmid bait coding for the first 548 amino-terminal residues of hSos1 yielded high numbers of positive, interacting clones. After successive rounds of screening, two clones showing positive interactions were selected for further study (Fig. 1c). After DNA sequencing, we found that in both cases the target cDNAs coded for the COP9 signalosome complex subunit 3 protein (CSN3) (NP_001303284.1). sos1-cops3-2 4 6 unspecified role unspecified role bait bait purified purified gst tag 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 region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association fig. 2a Cytoplasmic extracts from HeLa cells were incubated with purified GST-HD, GST-DH-PH, and GST-NDP fusion proteins coupled to glutathione–sepharose beads that were subsequently tested by means of immunoblotting with antibodies against CSN3 (Fig. 2a). Whereas purified GSTDHPH did not bind any CSN3, the GST-HD and GSTNDP peptides bound roughly similar amount of cellular CSN3 in lysates of actively growing HeLa cells (Fig. 2a, upper panel). sos1-cops3-3 1 7 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 6 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f2d Further confirmatory evidence supporting the in vivo interaction between Sos1 and the COP9 signalosome was obtained in similar, complementary assays using antibodies against CSN3 and Sos1 on lysates of cultured mouse NIH3T3 (not shown) or human HeLa cells (Fig. 2d), as well as of primary MEFs derived from WT or conditional, TAMinducible, Sos1-KO mice (Fig. 2e). cops3-sos1-1 3 8 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 5 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association fig.2e Further confirmatory evidence supporting the in vivo interaction between Sos1 and the COP9 signalosome was obtained in similar, complementary assays using antibodies against CSN3 and Sos1 on lysates of cultured mouse NIH3T3 (not shown) or human HeLa cells (Fig. 2d), as well as of primary MEFs derived from WT or conditional, TAMinducible, Sos1-KO mice (Fig. 2e).