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/ lassot-2001-3 ATF4 degradation relies on a phosphorylation-dependent interaction with the SCF(betaTrCP) ubiquitin ligase. 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 tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Lassot I., Ségéral E., Berlioz-Torrent C., Durand H., Groussin L., Hai T., Benarous R., Margottin-Goguet F. The identity of Skp1 was taken from PMID:9660940. benarous@cochin.inserm.fr Mol. Cell. Biol. (0270-7306) 2001 Only protein-protein interactions imex curation lassot-2001-1 ATF4 degradation relies on a phosphorylation-dependent interaction with the SCF(betaTrCP) ubiquitin ligase. baker's yeast-l40 Saccharomyces cerevisiae l40 l40 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 Lassot I., Ségéral E., Berlioz-Torrent C., Durand H., Groussin L., Hai T., Benarous R., Margottin-Goguet F. The identity of Skp1 was taken from PMID:9660940. benarous@cochin.inserm.fr Mol. Cell. Biol. (0270-7306) 2001 Only protein-protein interactions imex curation lassot-2001-2 ATF4 degradation relies on a phosphorylation-dependent interaction with the SCF(betaTrCP) ubiquitin ligase. 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 Lassot I., Ségéral E., Berlioz-Torrent C., Durand H., Groussin L., Hai T., Benarous R., Margottin-Goguet F. The identity of Skp1 was taken from PMID:9660940. benarous@cochin.inserm.fr human Jurkat cell cDNA library Mol. Cell. Biol. (0270-7306) 2001 Only protein-protein interactions imex curation lassot-2001-4 ATF4 degradation relies on a phosphorylation-dependent interaction with the SCF(betaTrCP) ubiquitin ligase. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lassot I., Ségéral E., Berlioz-Torrent C., Durand H., Groussin L., Hai T., Benarous R., Margottin-Goguet F. The identity of Skp1 was taken from PMID:9660940. benarous@cochin.inserm.fr Mol. Cell. Biol. (0270-7306) 2001 Only protein-protein interactions imex curation lassot-2001-5 ATF4 degradation relies on a phosphorylation-dependent interaction with the SCF(betaTrCP) ubiquitin ligase. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 confocal microscopy confocal microscopy anti tag immunost anti tag immunostaining anti tag immunost Lassot I., Ségéral E., Berlioz-Torrent C., Durand H., Groussin L., Hai T., Benarous R., Margottin-Goguet F. The identity of Skp1 was taken from PMID:9660940. benarous@cochin.inserm.fr Mol. Cell. Biol. (0270-7306) 2001 Only protein-protein interactions imex curation q9y297-2 F-box/WD repeat-containing protein 1A BTRC BTRCP FBW1A FBXW1A E3RSIkappaB Epididymis tissue protein Li 2a F-box and WD repeats protein beta-TrCP pIkappaBalpha-E3 receptor subunit protein protein human Homo sapiens Human MDPAEAVLQEKALKFMNSSEREDCNNGEPPRKIIPEKNSLRQTYNSCARLCLNQETVCLASTAMKTENCVAKTKLANGTSSMIVPKQRKLSASYEKEKELCVKYFEQWSESDQVEFVEHLISQMCHYQHGHINSYLKPMLQRDFITALPARGLDHIAENILSYLDAKSLCAAELVCKEWYRVTSDGMLWKKLIERMVRTDSLWRGLAERRGWGQYLFKNKPPDGNAPPNSFYRALYPKIIQDIETIESNWRCGRHSLQRIHCRSETSKGVYCLQYDDQKIVSGLRDNTIKIWDKNTLECKRILTGHTGSVLCLQYDERVIITGSSDSTVRVWDVNTGEMLNTLIHHCEAVLHLRFNNGMMVTCSKDRSIAVWDMASPTDITLRRVLVGHRAAVNVVDFDDKYIVSASGDRTIKVWNTSTCEFVRTLNGHKRGIACLQYRDRLVVSGSSDNTIRLWDIECGACLRVLEGHEELVRCIRFDNKRIVSGAYDGKIKVWDLVAALDPRAPAGTLCLRTLVEHSGRVFRLQFDEFQIVSSSHDDTILIWDFLNDPAAQAEPPRSPSRTYTYISR atf4_human Cyclic AMP-dependent transcription factor ATF-4 Activating transcription factor 4 CREB2 ATF4 TXREB Cyclic AMP-responsive element-binding protein 2 Tax-responsive enhancer element-binding protein 67 protein protein human Homo sapiens Human MTEMSFLSSEVLVGDLMSPFDQSGLGAEESLGLLDDYLEVAKHFKPHGFSSDKAKAGSSEWLAVDGLVSPSNNSKEDAFSGTDWMLEKMDLKEFDLDALLGIDDLETMPDDLLTTLDDTCDLFAPLVQETNKQPPQTVNPIGHLPESLTKPDQVAPFTFLQPLPLSPGVLSSTPDHSFSLELGSEVDITEGDRKPDYTAYVAMIPQCIKEEDTPSDNDSGICMSPESYLGSPQHSPSTRGSPNRSLPSPGVLCGSARPKPYDPPGEKMVAAKVKGEKLDKKLKKMEQNKTAATRYRQKKRAEQEALTGECKELEKKNEALKERADSLAKEIQYLKDLIEEVRKARGKKRVP 3BBB7379DC3B0D07 No Uniprot Update and Caution removed " The protein could not be merged with EBI-12907955 because of some conflicts with the protein sequence (features which cannot be shifted)." -EBI-12907955 no longer exists. ubiq_human UBIQ_HUMAN protein protein human Homo sapiens Human MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGG skp1_human S-phase kinase-associated protein 1 SKP1 EMC19 OCP2 SKP1A TCEB1L Cyclin-A/CDK2-associated protein p19 p19skp1 RNA polymerase II elongation factor-like protein Organ of Corti protein 2 Organ of Corti protein II SIII Transcription elongation factor B polypeptide 1-like protein protein human Homo sapiens Human MPSIKLQSSDGEIFEVDVEIAKQSVTIKTMLEDLGMDDEGDDDPVPLPNVNAAILKKVIQWCTHHKDDPPPPEDDENKEKRTDDIPVWDQEFLKVDQGTLFELILAANYLDIKGLLDVTCKTVANMIKGKTPEEIRKTFNIKNDFTEEEEAQVRKENQWCEEK o44382_drome slmb slimb CG3412 protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MMKMETDKIMDETNSNAQAFTTTMLYDPVRKKDSSPTYQTERELCFQYFTQWSESGQVDFVEHLLSRMCHYQHGQINAYLKPMLQRDFITLLPIKGLDHIGENILSYLDAESLKSSELVCKEWLRVISEGMLWKKLIERKVRTDSLWRGLAERRNWMQYLFKPRPGQTQRPHSFHRELFPKIMNDIDSIENNWRTGRHMLRRINCRSENSKGVYCLQYDDGKIVSGLRDNTIKIWDRTDLQCVKTLMGHTGSVLCLQYDDKVIISGSSDSTVRVWDVNTGEMVNTLIHHCEAVLHLRFNNGMMVTCSKDRSIAVWDMTSPSEITLRRVLVGHRAAVNVVDFDEKYIVSASGDRTIKVWSTSTCEFVRTLNGHKRGIACLQYRDRLVVSGSSDNSIRLWDIECGACLRVLEGHEELVRCIRFDTKRIVSGAYDGKIKVWDLVAALDPRAASNTLCLNTLVEHTGRVFRLQFDEFQIVSSSHDDTILIWDFLNFTPNENKTGRTPSPALMEH fbxw2_human F-box/WD repeat-containing protein 2 FBXW2 FBW2 FWD2 F-box and WD-40 domain-containing protein 2 Protein MD6 protein protein human Homo sapiens Human MERKDFETWLDNISVTFLSLTDLQKNETLDHLISLSGAVQLRHLSNNLETLLKRDFLKLLPLELSFYLLKWLDPQTLLTCCLVSKQWNKVISACTEVWQTACKNLGWQIDDSVQDALHWKKVYLKAILRMKQLEDHEAFETSSLIGHSARVYALYYKDGLLCTGSDDLSAKLWDVSTGQCVYGIQTHTCAAVKFDEQKLVTGSFDNTVACWEWSSGARTQHFRGHTGAVFSVDYNDELDILVSGSADFTVKVWALSAGTCLNTLTGHTEWVTKVVLQKCKVKSLLHSPGDYILLSADKYEIKIWPIGREINCKCLKTLSVSEDRSICLQPRLHFDGKYIVCSSALGLYQWDFASYDILRVIKTPEIANLALLGFGDIFALLFDNRYLYIMDLRTESLISRWPLPEYRKSKRGSSFLAGEASWLNGLDGHNDTGLVFATSMPDHSIHLVLWKEHG fbw1a_human F-box/WD repeat-containing protein 1A BTRC BTRCP FBW1A FBXW1A F-box and WD repeats protein beta-TrCP E3RSIkappaB pIkappaBalpha-E3 receptor subunit Epididymis tissue protein Li 2a protein protein human Homo sapiens Human MDPAEAVLQEKALKFMCSMPRSLWLGCSSLADSMPSLRCLYNPGTGALTAFQNSSEREDCNNGEPPRKIIPEKNSLRQTYNSCARLCLNQETVCLASTAMKTENCVAKTKLANGTSSMIVPKQRKLSASYEKEKELCVKYFEQWSESDQVEFVEHLISQMCHYQHGHINSYLKPMLQRDFITALPARGLDHIAENILSYLDAKSLCAAELVCKEWYRVTSDGMLWKKLIERMVRTDSLWRGLAERRGWGQYLFKNKPPDGNAPPNSFYRALYPKIIQDIETIESNWRCGRHSLQRIHCRSETSKGVYCLQYDDQKIVSGLRDNTIKIWDKNTLECKRILTGHTGSVLCLQYDERVIITGSSDSTVRVWDVNTGEMLNTLIHHCEAVLHLRFNNGMMVTCSKDRSIAVWDMASPTDITLRRVLVGHRAAVNVVDFDDKYIVSASGDRTIKVWNTSTCEFVRTLNGHKRGIACLQYRDRLVVSGSSDNTIRLWDIECGACLRVLEGHEELVRCIRFDNKRIVSGAYDGKIKVWDLVAALDPRAPAGTLCLRTLVEHSGRVFRLQFDEFQIVSSSHDDTILIWDFLNDPAAQAEPPRSPSRTYTYISR atf4-btrc-1 1450171 n/a 1450176 unspecified role unspecified role prey prey wd40 repeat required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1450177 unspecified role unspecified role bait bait P18848:p.Ser219Asn mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Fig. 2a, Fig. 2b, Fig. 2c Pretreatment of cells with 20 x 10E-6 molar Z-LLL-H/MG132 (N-benzyloxycarbonyl-L-leucyl-L-leucyl-L-leucinal) (CHEBI:75142), a proteasome inhibitor, increased the amount of precipitated Atf4, stabilizing the interaction with Btrc. btrc-atf4-2 1450171 n/a 1450177 unspecified role unspecified role prey prey n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1450176 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 physical association physical association false false false Fig. 3 ATF4-BTrCP interaction depends on ATF4 phosphorylation ubiq_human-atf4 1450171 n/a 1450178 unspecified role unspecified role bait bait n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1450177 unspecified role unspecified role prey prey n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Fig. 4 Expression of Btrc enhances ubiquitination of Atf4. Expression of the Btrc mutant lacking the F-box region, which anchors Btrc to the SCF ubiquitin ligase complex by binding to the Skp1 subunit, decreases ubiquitination of Atf4. btrc-atf4 1450172 n/a 1450176 unspecified role unspecified role bait bait wd40 repeat sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1450177 unspecified role unspecified role prey prey P18848:p.Ser219Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P18848:p.Asp218Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation 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 physical association physical association false false false Fig. 1b slmb-skp1 1450172 n/a 1450179 unspecified role unspecified role prey prey n/a 1450180 unspecified role unspecified role bait bait physical association physical association false false false Fig. 1c btrc-atf4-1 1450173 n/a 1450177 unspecified role unspecified role prey prey n/a 1450176 unspecified role unspecified role bait bait wd40 repeat sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false fbxw2-skp1-1 1450173 n/a 1450181 unspecified role unspecified role bait bait n/a 1450179 unspecified role unspecified role prey prey physical association physical association false false false Fig. 1c atf4-btrc-2 1450174 n/a 1450182 unspecified role unspecified role prey prey n/a 1450177 unspecified role unspecified role bait bait physical association physical association false false false Fig. 2e atf4-btrc-3 1450175 n/a 1450176 tag fluorescence tag visualisation by fluorescence tag fluorescence unspecified role unspecified role neutral component neutral component c-terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1450177 unspecified role unspecified role neutral component neutral component n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false colocalization colocalization false false false Fig. 5c