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 TRIAD1 and HHARI bind to and are activated by distinct neddylated Cullin-RING ligase complexes. TRIAD1 and HHARI bind to and are activated by distinct neddylated Cullin-RING ligase complexes. EMBO J. (0261-4189) 2013 imex curation Kelsall I.R., Duda D.M., Olszewski J.L., Hofmann K., Knebel A., Langevin F., Wood N., Wightman M., Schulman B.A., Alpi A.F. a.f.alpi@dundee.ac.uk 18-Sep-2014: Contacted by IntAct-Help. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip ms participant Identification by mass spectrometry ms participant Kelsall I.R., Duda D.M., Olszewski J.L., Hofmann K., Knebel A., Langevin F., Wood N., Wightman M., Schulman B.A., Alpi A.F. a.f.alpi@dundee.ac.uk Accepted 2014-SEP-12 by MARTA EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation TRIAD1 and HHARI bind to and are activated by distinct neddylated Cullin-RING ligase complexes. TRIAD1 and HHARI bind to and are activated by distinct neddylated Cullin-RING ligase complexes. EMBO J. (0261-4189) 2013 imex curation Kelsall I.R., Duda D.M., Olszewski J.L., Hofmann K., Knebel A., Langevin F., Wood N., Wightman M., Schulman B.A., Alpi A.F. a.f.alpi@dundee.ac.uk 18-Sep-2014: Contacted by IntAct-Help. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Kelsall I.R., Duda D.M., Olszewski J.L., Hofmann K., Knebel A., Langevin F., Wood N., Wightman M., Schulman B.A., Alpi A.F. a.f.alpi@dundee.ac.uk Accepted 2014-SEP-12 by MARTA EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation TRIAD1 and HHARI bind to and are activated by distinct neddylated Cullin-RING ligase complexes. TRIAD1 and HHARI bind to and are activated by distinct neddylated Cullin-RING ligase complexes. EMBO J. (0261-4189) 2013 imex curation Kelsall I.R., Duda D.M., Olszewski J.L., Hofmann K., Knebel A., Langevin F., Wood N., Wightman M., Schulman B.A., Alpi A.F. a.f.alpi@dundee.ac.uk 18-Sep-2014: Contacted by IntAct-Help. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Kelsall I.R., Duda D.M., Olszewski J.L., Hofmann K., Knebel A., Langevin F., Wood N., Wightman M., Schulman B.A., Alpi A.F. a.f.alpi@dundee.ac.uk Accepted 2014-SEP-12 by MARTA EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation cul5_human Cullin-5 CUL5 VACM1 Vasopressin-activated calcium-mobilizing receptor 1 protein protein human Homo sapiens Human MATSNLLKNKGSLQFEDKWDFMRPIVLKLLRQESVTKQQWFDLFSDVHAVCLWDDKGPAKIHQALKEDILEFIKQAQARVLSHQDDTALLKAYIVEWRKFFTQCDILPKPFCQLEITLMGKQGSNKKSNVEDSIVRKLMLDTWNESIFSNIKNRLQDSAMKLVHAERLGEAFDSQLVIGVRESYVNLCSNPEDKLQIYRDNFEKAYLDSTERFYRTQAPSYLQQNGVQNYMKYADAKLKEEEKRALRYLETRRECNSVEALMECCVNALVTSFKETILAECQGMIKRNETEKLHLMFSLMDKVPNGIEPMLKDLEEHIISAGLADMVAAAETITTDSEKYVEQLLTLFNRFSKLVKEAFQDDPRFLTARDKAYKAVVNDATIFKLELPLKQKGVGLKTQPESKCPELLANYCDMLLRKTPLSKKLTSEEIEAKLKEVLLVLKYVQNKDVFMRYHKAHLTRRLILDISADSEIEENMVEWLREVGMPADYVNKLARMFQDIKVSEDLNQAFKEMHKNNKLALPADSVNIKILNAGAWSRSSEKVFVSLPTELEDLIPEVEEFYKKNHSGRKLHWHHLMSNGIITFKNEVGQYDLEVTTFQLAVLFAWNQRPREKISFENLKLATELPDAELRRTLWSLVAFPKLKRQVLLYEPQVNSPKDFTEGTLFSVNQEFSLIKNAKVQKRGKINLIGRLQLTTERMREEENEGIVQLRILRTQEAIIQIMKMRKKISNAQLQTELVEILKNMFLPQKKMIKEQIEWLIEHKYIRRDESDINTFIYMA DIP protein Q93034 original sequence version: 160 DIP protein Q93034 original sequence version: 128 DIP protein Q93034 original sequence version: 126 DIP protein Q93034 original sequence version: 142 DIP protein Q93034 original sequence version: 96 57463CB4ED76E303 ari2_human E3 ubiquitin-protein ligase ARIH2 Triad1 protein ARIH2 ARI2 TRIAD1 HT005 RING-type E3 ubiquitin transferase ARIH2 protein protein human Homo sapiens Human MSVDMNSQGSDSNEEDYDPNCEEEEEEEEDDPGDIEDYYVGVASDVEQQGADAFDPEEYQFTCLTYKESEGALNEHMTSLASVLKVSHSVAKLILVNFHWQVSEILDRYKSNSAQLLVEARVQPNPSKHVPTSHPPHHCAVCMQFVRKENLLSLACQHQFCRSCWEQHCSVLVKDGVGVGVSCMAQDCPLRTPEDFVFPLLPNEELREKYRRYLFRDYVESHYQLQLCPGADCPMVIRVQEPRARRVQCNRCNEVFCFKCRQMYHAPTDCATIRKWLTKCADDSETANYISAHTKDCPKCNICIEKNGGCNHMQCSKCKHDFCWMCLGDWKTHGSEYYECSRYKENPDIVNQSQQAQAREALKKYLFYFERWENHNKSLQLEAQTYQRIHEKIQERVMNNLGTWIDWQYLQNAAKLLAKCRYTLQYTYPYAYYMESGPRKKLFEYQQAQLEAEIENLSWKVERADSYDRGDLENQMHIAEQRRRTLLKDFHDT 30AFFDD327B51013 cul1_human Cullin-1 CUL1 protein protein human Homo sapiens Human MSSTRSQNPHGLKQIGLDQIWDDLRAGIQQVYTRQSMAKSRYMELYTHVYNYCTSVHQSNQARGAGVPPSKSKKGQTPGGAQFVGLELYKRLKEFLKNYLTNLLKDGEDLMDESVLKFYTQQWEDYRFSSKVLNGICAYLNRHWVRRECDEGRKGIYEIYSLALVTWRDCLFRPLNKQVTNAVLKLIEKERNGETINTRLISGVVQSYVELGLNEDDAFAKGPTLTVYKESFESQFLADTERFYTRESTEFLQQNPVTEYMKKAEARLLEEQRRVQVYLHESTQDELARKCEQVLIEKHLEIFHTEFQNLLDADKNEDLGRMYNLVSRIQDGLGELKKLLETHIHNQGLAAIEKCGEAALNDPKMYVQTVLDVHKKYNALVMSAFNNDAGFVAALDKACGRFINNNAVTKMAQSSSKSPELLARYCDSLLKKSSKNPEEAELEDTLNQVMVVFKYIEDKDVFQKFYAKMLAKRLVHQNSASDDAEASMISKLKQACGFEYTSKLQRMFQDIGVSKDLNEQFKKHLTNSEPLDLDFSIQVLSSGSWPFQQSCTFALPSELERSYQRFTAFYASRHSGRKLTWLYQLSKGELVTNCFKNRYTLQASTFQMAILLQYNTEDAYTVQQLTDSTQIKMDILAQVLQILLKSKLLVLEDENANVDEVELKPDTLIKLYLGYKNKKLRVNINVPMKTEQKQEQETTHKNIEEDRKLLIQAAIVRIMKMRKVLKHQQLLGEVLTQLSSRFKPRVPVIKKCIDILIEKEYLERVDGEKDTYSYLA DIP protein Q13616 original sequence version: 144 DIP protein Q13616 original sequence version: 133 DIP protein Q13616 original sequence version: null DIP protein Q13616 original sequence version: 135 DIP protein Q13616 original sequence version: 98 DIP protein Q13616 original sequence version: 110 DIP protein Q13616 original sequence version: 146 DIP protein Q13616 original sequence version: 148 DIP protein Q13616 original sequence version: 100 DIP protein Q13616 original sequence version: 85 DIP protein Q13616 original sequence version: 114 DIP protein Q13616 original sequence version: 119 DIP protein Q13616 original sequence version: 136 DIP protein Q13616 original sequence version: 117 DIP protein Q13616 original sequence version: 99 DIP protein Q13616 original sequence version: 108 6625A1FFA7799BBA cul2_human Cullin-2 CUL2 protein protein human Homo sapiens Human MSLKPRVVDFDETWNKLLTTIKAVVMLEYVERATWNDRFSDIYALCVAYPEPLGERLYTETKIFLENHVRHLHKRVLESEEQVLVMYHRYWEEYSKGADYMDCLYRYLNTQFIKKNKLTEADLQYGYGGVDMNEPLMEIGELALDMWRKLMVEPLQAILIRMLLREIKNDRGGEDPNQKVIHGVINSFVHVEQYKKKFPLKFYQEIFESPFLTETGEYYKQEASNLLQESNCSQYMEKVLGRLKDEEIRCRKYLHPSSYTKVIHECQQRMVADHLQFLHAECHNIIRQEKKNDMANMYVLLRAVSTGLPHMIQELQNHIHDEGLRATSNLTQENMPTLFVESVLEVHGKFVQLINTVLNGDQHFMSALDKALTSVVNYREPKSVCKAPELLAKYCDNLLKKSAKGMTENEVEDRLTSFITVFKYIDDKDVFQKFYARMLAKRLIHGLSMSMDSEEAMINKLKQACGYEFTSKLHRMYTDMSVSADLNNKFNNFIKNQDTVIDLGISFQIYVLQAGAWPLTQAPSSTFAIPQELEKSVQMFELFYSQHFSGRKLTWLHYLCTGEVKMNYLGKPYVAMVTTYQMAVLLAFNNSETVSYKELQDSTQMNEKELTKTIKSLLDVKMINHDSEKEDIDAESSFSLNMNFSSKRTKFKITTSMQKDTPQEMEQTRSAVDEDRKMYLQAAIVRIMKARKVLRHNALIQEVISQSRARFNPSISMIKKCIEVLIDKQYIERSQASADEYSYVA DIP protein Q13617 original sequence version: 129 DIP protein Q13617 original sequence version: 127 DIP protein Q13617-1 original sequence version: 139 DIP protein Q13617 original sequence version: 111 DIP protein Q13617 original sequence version: 102 30647248F671AB0E eloc_human Elongin-C ELOC Elongin 15 kDa subunit RNA polymerase II transcription factor SIII subunit C SIII p15 TCEB1 Transcription elongation factor B polypeptide 1 protein protein human Homo sapiens Human MDGEEKTYGGCEGPDAMYVKLISSDGHEFIVKREHALTSGTIKAMLSGPGQFAENETNEVNFREIPSHVLSKVCMYFTYKVRYTNSSTEIPEFPIAPEIALELLMAANFLDC 98D88696E883538B cul3_human Cullin-3 CUL3 KIAA0617 protein protein human Homo sapiens Human MSNLSKGTGSRKDTKMRIRAFPMTMDEKYVNSIWDLLKNAIQEIQRKNNSGLSFEELYRNAYTMVLHKHGEKLYTGLREVVTEHLINKVREDVLNSLNNNFLQTLNQAWNDHQTAMVMIRDILMYMDRVYVQQNNVENVYNLGLIIFRDQVVRYGCIRDHLRQTLLDMIARERKGEVVDRGAIRNACQMLMILGLEGRSVYEEDFEAPFLEMSAEFFQMESQKFLAENSASVYIKKVEARINEEIERVMHCLDKSTEEPIVKVVERELISKHMKTIVEMENSGLVHMLKNGKTEDLGCMYKLFSRVPNGLKTMCECMSSYLREQGKALVSEEGEGKNPVDYIQGLLDLKSRFDRFLLESFNNDRLFKQTIAGDFEYFLNLNSRSPEYLSLFIDDKLKKGVKGLTEQEVETILDKAMVLFRFMQEKDVFERYYKQHLARRLLTNKSVSDDSEKNMISKLKTECGCQFTSKLEGMFRDMSISNTTMDEFRQHLQATGVSLGGVDLTVRVLTTGYWPTQSATPKCNIPPAPRHAFEIFRRFYLAKHSGRQLTLQHHMGSADLNATFYGPVKKEDGSEVGVGGAQVTGSNTRKHILQVSTFQMTILMLFNNREKYTFEEIQQETDIPERELVRALQSLACGKPTQRVLTKEPKSKEIENGHIFTVNDQFTSKLHRVKIQTVAAKQGESDPERKETRQKVDDDRKHEIEAAIVRIMKSRKKMQHNVLVAEVTQQLKARFLPSPVVIKKRIEGLIEREYLARTPEDRKVYTYVA DIP protein Q13618 original sequence version: 140 DIP protein Q13618 original sequence version: 163 DIP protein Q13618 original sequence version: 121 DIP protein Q13618 original sequence version: 161 A1A02022480BF099 cul4a_human Cullin-4A CUL4A protein protein human Homo sapiens Human MADEAPRKGSFSALVGRTNGLTKPAALAAAPAKPGGAGGSKKLVIKNFRDRPRLPDNYTQDTWRKLHEAVRAVQSSTSIRYNLEELYQAVENLCSHKVSPMLYKQLRQACEDHVQAQILPFREDSLDSVLFLKKINTCWQDHCRQMIMIRSIFLFLDRTYVLQNSTLPSIWDMGLELFRTHIISDKMVQSKTIDGILLLIERERSGEAVDRSLLRSLLGMLSDLQVYKDSFELKFLEETNCLYAAEGQRLMQEREVPEYLNHVSKRLEEEGDRVITYLDHSTQKPLIACVEKQLLGEHLTAILQKGLDHLLDENRVPDLAQMYQLFSRVRGGQQALLQHWSEYIKTFGTAIVINPEKDKDMVQDLLDFKDKVDHVIEVCFQKNERFVNLMKESFETFINKRPNKPAELIAKHVDSKLRAGNKEATDEELERTLDKIMILFRFIHGKDVFEAFYKKDLAKRLLVGKSASVDAEKSMLSKLKHECGAAFTSKLEGMFKDMELSKDIMVHFKQHMQNQSDSGPIDLTVNILTMGYWPTYTPMEVHLTPEMIKLQEVFKAFYLGKHSGRKLQWQTTLGHAVLKAEFKEGKKEFQVSLFQTLVLLMFNEGDGFSFEEIKMATGIEDSELRRTLQSLACGKARVLIKSPKGKEVEDGDKFIFNGEFKHKLFRIKINQIQMKETVEEQVSTTERVFQDRQYQIDAAIVRIMKMRKTLGHNLLVSELYNQLKFPVKPGDLKKRIESLIDRDYMERDKDNPNQYHYVA DIP protein Q13619 original sequence version: 136 DIP protein Q13619 original sequence version: 131 DIP protein Q13619 original sequence version: 129 DIP protein Q13619 original sequence version: 87 DIP protein Q13619 original sequence version: 104 DIP protein Q13619 original sequence version: 151 DIP protein Q13619 original sequence version: 111 DIP protein Q13619 original sequence version: 140 3C4C6A1BBD94D51B elob_human Elongin-B ELOB Elongin 18 kDa subunit RNA polymerase II transcription factor SIII subunit B SIII p18 TCEB2 Transcription elongation factor B polypeptide 2 protein protein human Homo sapiens Human MDVFLMIRRHKTTIFTDAKESSTVFELKRIVEGILKRPPDEQRLYKDDQLLDDGKTLGECGFTSQTARPQAPATVGLAFRADDTFEALCIEPFSSPPELPDVMKPQDSGSSANEQAVQ C045F58FBED0EC47 cdn1b_human Cyclin-dependent kinase inhibitor 1B Cyclin-dependent kinase inhibitor p27 p27Kip1 CDKN1B KIP1 protein protein human Homo sapiens Human MSNVRVSNGSPSLERMDARQAEHPKPSACRNLFGPVDHEELTRDLEKHCRDMEEASQRKWNFDFQNHKPLEGKYEWQEVEKGSLPEFYYRPPRPPKGACKVPAQESQDVSGSRPAAPLIGAPANSEDTHLVDPKTDPSDSQTGLAEQCAGIRKRPATDDSSTQNKRANRTEENVSDGSPNAGSVEQTPKKPGLRRRQT DIP protein P46527 original sequence version: 120 DIP protein P46527 original sequence version: 115 DIP protein P46527 original sequence version: 154 DIP protein P46527 original sequence version: 168 DIP protein P46527 original sequence version: 164 DIP protein P46527 original sequence version: 118 1118D58901CDF3FC ub2l3_human Ubiquitin-conjugating enzyme E2 L3 Ubiquitin-protein ligase L3 Ubiquitin carrier protein L3 UbcH7 Ubiquitin-conjugating enzyme E2-F1 L-UBC UBE2L3 UBCE7 UBCH7 E2 ubiquitin-conjugating enzyme L3 protein protein human Homo sapiens Human MAASRRLMKELEEIRKCGMKNFRNIQVDEANLLTWQGLIVPDNPPYDKGAFRIEINFPAEYPFKPPKITFKTKIYHPNIDEKGQVCLPVISAENWKPATKTDQVIQSLIALVNDPQPEHPLRADLAEEYSKDRKKFCKNAEEFTKKYGEKRPVD F5A30243BE3C9985 ccne1_human G1/S-specific cyclin-E1 CCNE1 CCNE protein protein human Homo sapiens Human MPRERRERDAKERDTMKEDGGAEFSARSRKRKANVTVFLQDPDEEMAKIDRTARDQCGSQPWDNNAVCADPCSLIPTPDKEDDDRVYPNSTCKPRIIAPSRGSPLPVLSWANREEVWKIMLNKEKTYLRDQHFLEQHPLLQPKMRAILLDWLMEVCEVYKLHRETFYLAQDFFDRYMATQENVVKTLLQLIGISSLFIAAKLEEIYPPKLHQFAYVTDGACSGDEILTMELMIMKALKWRLSPLTIVSWLNVYMQVAYLNDLHEVLLPQYPQQIFIQIAELLDLCVLDVDCLEFPYGILAASALYHFSSSELMQKVSGYQWCDIENCVKWMVPFAMVIRETGSSKLKHFRGVADEDAHNIQTHRDSLDLLDKARAKKAMLSEQNRASPLPSGLLTPPQSGKKQSSGPEMA DIP protein P24864 original sequence version: 114 DIP protein P24864 original sequence version: 150 424DF0B253B7047E ari1_human E3 ubiquitin-protein ligase ARIH1 Protein ariadne-1 homolog ARIH1 ARI MOP6 UBCH7BP Ubiquitin-conjugating enzyme E2-binding protein 1 UbcH7-binding protein UbcM4-interacting protein HHARI H7-AP2 Monocyte protein 6 HUSSY-27 protein protein human Homo sapiens Human MDSDEGYNYEFDEDEECSEEDSGAEEEEDEDDDEPDDDTLDLGEVELVEPGLGVGGERDGLLCGETGGGGGSALGPGGGGGGGGGGGGGGPGHEQEEDYRYEVLTAEQILQHMVECIREVNEVIQNPATITRILLSHFNWDKEKLMERYFDGNLEKLFAECHVINPSKKSRTRQMNTRSSAQDMPCQICYLNYPNSYFTGLECGHKFCMQCWSEYLTTKIMEEGMGQTISCPAHGCDILVDDNTVMRLITDSKVKLKYQHLITNSFVECNRLLKWCPAPDCHHVVKVQYPDAKPVRCKCGRQFCFNCGENWHDPVKCKWLKKWIKKCDDDSETSNWIAANTKECPKCHVTIEKDGGCNHMVCRNQNCKAEFCWVCLGPWEPHGSAWYNCNRYNEDDAKAARDAQERSRAALQRYLFYCNRYMNHMQSLRFEHKLYAQVKQKMEEMQQHNMSWIEVQFLKKAVDVLCQCRATLMYTYVFAFYLKKNNQSIIFENNQADLENATEVLSGYLERDISQDSLQDIKQKVQDKYRYCESRRRVLLQHVHEGYEKDLWEYIED DIP protein Q9Y4X5 original sequence version: 122 DIP protein Q9Y4X5 original sequence version: 103 DFFF8965DAB41DC8 arih2-ube2l3-2 3 13 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 5 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f1a arih2-ube2l3-3 2 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell O95376:p.His158Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain H A Wed May 02 16:56:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his158ala' Tue Jun 30 19:44:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His158Ala' tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f1c arih2-cul5 1 4 ms participant Identification by mass spectrometry ms participant ms participant Identification by mass spectrometry ms participant unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 13 ms participant Identification by mass spectrometry ms participant ms participant Identification by mass spectrometry ms participant unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 5 ms participant Identification by mass spectrometry ms participant ms participant Identification by mass spectrometry ms participant unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association f2a arih2-ube2l3-4 3 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 5 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f2b cul5-arih2 3 5 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f2b arih2-cul5-1 2 13 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined range sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association f2d sf2b f7a arih2-cul5-2 2 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell range sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f2d sf2b arih2-cul5-4 2 11 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 8 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association f3a arih2-cul5-5 2 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell O95376:p.[Leu84Ala;Val86Ala;Trp100Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain V A certain certain sequence position certain certain certain sequence position certain L A certain certain sequence position certain certain certain sequence position certain W A Fri Jul 22 11:12:12 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'val86ala,leu84ala,trp100ala' Wed May 02 17:31:01 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu84ala,val86ala,trp100ala' Tue Jun 30 19:24:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu84Ala;Val86Ala;Trp100Ala]' tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f3e arih2-cul5-6 2 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell O95376:p.Trp100Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain W A Wed May 02 19:08:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp100ala' Tue Jun 30 16:00:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp100Ala' tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f3e arih2-cul5-7 2 15 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 13 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association f5a f5b arih1-cul2 2 9 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 10 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 15 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 13 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association sf4 Strikingly, GFP‐HHARI interacted with the neddylated form of CUL1, ‐2, ‐3 and ‐4A but not with CUL4B or CUL5 (Figure 5A; Supplementary Figure S4). arih1-cul1 2 13 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 15 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q9Y4X5:p.His205Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain H A Wed May 02 16:07:21 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his205ala' Tue Jun 30 17:22:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His205Ala' association association f5b Precipitation of GFP‐HHARI with either an N‐terminal 100‐aa deletion (GFP‐HHARI (aa 101–557)) or triple mutations in the predicted UBA‐like domain (V123A, I124A, and W140A) abolished or significantly reduced the interaction with CUL1‐NEDD8 (Figure 5B and C), whereas the RING1 H205A mutation significantly decreased UBCH7 binding without disrupting the interaction with CUL1‐NEDD8. arih1-cul1-1 2 15 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined range binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Q9Y4X5:p.[Val123Ala;Ile124Ala;Trp140Ala] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain V A certain certain sequence position certain certain certain sequence position certain I A certain certain sequence position certain certain certain sequence position certain W A Wed May 02 15:40:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val123ala,ile124ala,trp140ala' Tue Jun 30 18:47:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Val123Ala;Ile124Ala;Trp140Ala]' 6 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f5c f5b Precipitation of GFP‐HHARI with either an N‐terminal 100‐aa deletion (GFP‐HHARI (aa 101–557)) or triple mutations in the predicted UBA‐like domain (V123A, I124A, and W140A) abolished or significantly reduced the interaction with CUL1‐NEDD8 (Figure 5B and C), whereas the RING1 H205A mutation significantly decreased UBCH7 binding without disrupting the interaction with CUL1‐NEDD8. arih1-cul1-2 2 15 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell Q9Y4X5:p.Val123Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain V A Wed May 02 19:07:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val123ala' Tue Jun 30 18:49:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val123Ala' tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined range binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 6 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f5c f5b arih1-cul1-3 2 6 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 13 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 15 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell Q9Y4X5:p.[Phe430Ala;Glu431Ala;Glu503Ala] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain F A certain certain sequence position certain certain certain sequence position certain E A certain certain sequence position certain certain certain sequence position certain E A Wed May 02 17:35:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe430ala,glu431ala,glu503ala' Tue Jun 30 19:50:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe430Ala;Glu431Ala;Glu503Ala]' tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association sf5f We next tested the effects of mutations in the HHARI Ariadne domain, which were recently shown to relieve auto‐inhibition and result in enhanced HHARI auto‐ubiquitylation in vitro (Duda et al, 2013) (Figure 7H). We observed less GFP‐HHARI overall, potentially reflecting effects of auto‐ubiquitylation, and a concomitant increase in CUL1‐NEDD8 levels. Inactivation of E3 ligase activity by means of an additional catalytic C357S mutation restored accumulation of these Ariadne domain variants, as did treatment with the proteasomal inhibitor MG132 (Supplementary Figure S5F). arih2-cul5-9 2 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined O95376:p.Cys310Ser mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain C S Wed May 02 18:38:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys310ser' Tue Jun 30 18:21:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys310Ser' 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f7b arih2-cul5-10 2 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined O95376:p.Cys318Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain C A Wed May 02 19:57:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys318ala' Tue Jun 30 16:21:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys318Ala' physical association physical association f7b arih2-cul5-11 2 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell O95376:p.Cys310Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain C A Wed May 02 16:32:48 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys310ala' Tue Jun 30 18:51:51 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys310Ala' tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f7b arih2-cul5-12 2 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined O95376:p.Cys300Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain C A Wed May 02 18:42:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys300ala' Tue Jun 30 17:18:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys300Ala' 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f7b f7a arih1-cul1-4 2 6 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 15 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q9Y4X5:p.Cys375Ser mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain C S Wed May 02 16:33:48 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys375ser' Tue Jun 30 20:00:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys375Ser' physical association physical association f7d arih1-ccne1 2 14 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 12 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 6 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 15 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q9Y4X5:p.Cys357Ser mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain C S Wed May 02 19:41:47 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys357ser' Tue Jun 30 17:26:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys357Ser' 13 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association f7h We next tested the effects of mutations in the HHARI Ariadne domain, which were recently shown to relieve auto‐inhibition and result in enhanced HHARI auto‐ubiquitylation in vitro (Duda et al, 2013) (Figure 7H). We observed less GFP‐HHARI overall, potentially reflecting effects of auto‐ubiquitylation, and a concomitant increase in CUL1‐NEDD8 levels.