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/ yokoyama-2010-8 A higher-order complex containing AF4 and ENL family proteins with P-TEFb facilitates oncogenic and physiologic MLL-dependent transcription. 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 tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Yokoyama A., Lin M., Naresh A., Kitabayashi I., Cleary ML. Apoptosis - Interactions involving proteins with a function related to apoptosis Anti AF4 monoclonal antibodies were X linked to G Sepharose beads. Additional detail of constructs supplied by the author. ayokoyam@ncc.go.jp, mcleary@stanford.edu Cancer Cell (1535-6108) 2010 Only protein-protein interactions 31 imex curation Cancer - Interactions investigated in the context of cancer yokoyama-2010-3 A higher-order complex containing AF4 and ENL family proteins with P-TEFb facilitates oncogenic and physiologic MLL-dependent transcription. human-k_562 Homo sapiens myeloid leukemia cells. k562 Myelogenous leukemia K562 cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Yokoyama A., Lin M., Naresh A., Kitabayashi I., Cleary ML. Apoptosis - Interactions involving proteins with a function related to apoptosis Anti AF4 monoclonal antibodies were X linked to G Sepharose beads. Additional detail of constructs supplied by the author. ayokoyam@ncc.go.jp, mcleary@stanford.edu Cancer Cell (1535-6108) 2010 Only protein-protein interactions 31 imex curation Cancer - Interactions investigated in the context of cancer yokoyama-2010-7 A higher-order complex containing AF4 and ENL family proteins with P-TEFb facilitates oncogenic and physiologic MLL-dependent transcription. 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 tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Yokoyama A., Lin M., Naresh A., Kitabayashi I., Cleary ML. Apoptosis - Interactions involving proteins with a function related to apoptosis Anti AF4 monoclonal antibodies were X linked to G Sepharose beads. Additional detail of constructs supplied by the author. ayokoyam@ncc.go.jp, mcleary@stanford.edu Cancer Cell (1535-6108) 2010 Only protein-protein interactions 31 imex curation Cancer - Interactions investigated in the context of cancer yokoyama-2010-4 A higher-order complex containing AF4 and ENL family proteins with P-TEFb facilitates oncogenic and physiologic MLL-dependent transcription. human-hb1119 Homo sapiens acute lymphoblastic leukemia cell line HB1119 hb1119 Human acute lymphoblastic leukemia cell line HB1119 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Yokoyama A., Lin M., Naresh A., Kitabayashi I., Cleary ML. Apoptosis - Interactions involving proteins with a function related to apoptosis Trembl entries for fusion proteins are given as a reference only. Anti AF4 monoclonal antibodies were X linked to G Sepharose beads. Additional detail of constructs supplied by the author. ayokoyam@ncc.go.jp, mcleary@stanford.edu Cancer Cell (1535-6108) 2010 Only protein-protein interactions 31 imex curation Cancer - Interactions investigated in the context of cancer yokoyama-2010-5 A higher-order complex containing AF4 and ENL family proteins with P-TEFb facilitates oncogenic and physiologic MLL-dependent transcription. human-kp-l-ry Human: KP-L-RY leukemic blast cell line kp-l-ry Human KP-L-RY leukemic blast cell line Lymphoblast cell line obtained from the peripheral blood of an ALL infant with MLL rearrangement. In the fusion transcript the MLL exon 6 is fused in frame to the 5' side of the putative transactivation domain of AF5q31. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Yokoyama A., Lin M., Naresh A., Kitabayashi I., Cleary ML. Apoptosis - Interactions involving proteins with a function related to apoptosis Trembl entries for fusion proteins are given as a reference only. Anti AF4 monoclonal antibodies were X linked to G Sepharose beads. Additional detail of constructs supplied by the author. ayokoyam@ncc.go.jp, mcleary@stanford.edu Cancer Cell (1535-6108) 2010 Only protein-protein interactions 31 imex curation Cancer - Interactions investigated in the context of cancer yokoyama-2010-6 A higher-order complex containing AF4 and ENL family proteins with P-TEFb facilitates oncogenic and physiologic MLL-dependent transcription. human-ml_2 Homo sapiens ML-2 acute myeloid leukemia-derived cell line ml_2 Human acute myeloid leukemia-derived cell line ML-2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Yokoyama A., Lin M., Naresh A., Kitabayashi I., Cleary ML. Apoptosis - Interactions involving proteins with a function related to apoptosis Anti AF4 monoclonal antibodies were X linked to G Sepharose beads. Additional detail of constructs supplied by the author. ayokoyam@ncc.go.jp, mcleary@stanford.edu Cancer Cell (1535-6108) 2010 Only protein-protein interactions 31 imex curation Cancer - Interactions investigated in the context of cancer yokoyama-2010-9 A higher-order complex containing AF4 and ENL family proteins with P-TEFb facilitates oncogenic and physiologic MLL-dependent transcription. human-sem Human acute lymphoblastic leukemia cell line SEM sem Human acute lymphoblastic leukemia cell line SEM SEM-K2 cells express amplified wild-type FLT3 along with the MLL-AF4 fusion protein. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Yokoyama A., Lin M., Naresh A., Kitabayashi I., Cleary ML. Apoptosis - Interactions involving proteins with a function related to apoptosis Anti AF4 monoclonal antibodies were X linked to G Sepharose beads. Cell line used was SEM-K2 -a subclone of the SEM cell line. Additional detail of constructs supplied by the author. ayokoyam@ncc.go.jp, mcleary@stanford.edu Cancer Cell (1535-6108) 2010 Only protein-protein interactions 31 imex curation Cancer - Interactions investigated in the context of cancer Anti AF4 monoclonal antibodies were X linked to G Sepharose beads. yokoyama-2010-1 A higher-order complex containing AF4 and ENL family proteins with P-TEFb facilitates oncogenic and physiologic MLL-dependent transcription. human-k_562 Homo sapiens myeloid leukemia cells. k562 Myelogenous leukemia K562 cells affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom ms participant Identification by mass spectrometry ms participant Yokoyama A., Lin M., Naresh A., Kitabayashi I., Cleary ML. Apoptosis - Interactions involving proteins with a function related to apoptosis Anti AF4 monoclonal antibodies were X linked to G Sepharose beads. Additional detail of constructs supplied by the author. ayokoyam@ncc.go.jp, mcleary@stanford.edu Cancer Cell (1535-6108) 2010 Only protein-protein interactions 31 imex curation Cancer - Interactions investigated in the context of cancer yokoyama-2010-2 A higher-order complex containing AF4 and ENL family proteins with P-TEFb facilitates oncogenic and physiologic MLL-dependent transcription. human-ml_2 Homo sapiens ML-2 acute myeloid leukemia-derived cell line ml_2 Human acute myeloid leukemia-derived cell line ML-2 affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom ms participant Identification by mass spectrometry ms participant Yokoyama A., Lin M., Naresh A., Kitabayashi I., Cleary ML. Apoptosis - Interactions involving proteins with a function related to apoptosis Anti AF4 monoclonal antibodies were X linked to G Sepharose beads. Additional detail of constructs supplied by the author. ayokoyam@ncc.go.jp, mcleary@stanford.edu Cancer Cell (1535-6108) 2010 Only protein-protein interactions 31 imex curation Cancer - Interactions investigated in the context of cancer dot1l_human Histone-lysine N-methyltransferase, H3 lysine-79 specific DOT1L KIAA1814 KMT4 Histone H3-K79 methyltransferase DOT1-like protein Lysine N-methyltransferase 4 protein protein human Homo sapiens Human MGEKLELRLKSPVGAEPAVYPWPLPVYDKHHDAAHEIIETIRWVCEEIPDLKLAMENYVLIDYDTKSFESMQRLCDKYNRAIDSIHQLWKGTTQPMKLNTRPSTGLLRHILQQVYNHSVTDPEKLNNYEPFSPEVYGETSFDLVAQMIDEIKMTDDDLFVDLGSGVGQVVLQVAAATNCKHHYGVEKADIPAKYAETMDREFRKWMKWYGKKHAEYTLERGDFLSEEWRERIANTSVIFVNNFAFGPEVDHQLKERFANMKEGGRIVSSKPFAPLNFRINSRNLSDIGTIMRVVELSPLKGSVSWTGKPVSYYLHTIDRTILENYFSSLKNPKLREEQEAARRRQQRESKSNAATPTKGPEGKVAGPADAPMDSGAEEEKAGAATVKKPSPSKARKKKLNKKGRKMAGRKRGRPKKMNTANPERKPKKNQTALDALHAQTVSQTAASSPQDAYRSPHSPFYQLPPSVQRHSPNPLLVAPTPPALQKLLESFKIQYLQFLAYTKTPQYKASLQELLGQEKEKNAQLLGAAQQLLSHCQAQKEEIRRLFQQKLDELGVKALTYNDLIQAQKEISAHNQQLREQSEQLEQDNRALRGQSLQLLKARCEELQLDWATLSLEKLLKEKQALKSQISEKQRHCLELQISIVELEKSQRQQELLQLKSCVPPDDALSLHLRGKGALGRELEPDASRLHLELDCTKFSLPHLSSMSPELSMNGQAAGYELCGVLSRPSSKQNTPQYLASPLDQEVVPCTPSHVGRPRLEKLSGLAAPDYTRLSPAKIVLRRHLSQDHTVPGRPAASELHSRAEHTKENGLPYQSPSVPGSMKLSPQDPRPLSPGALQLAGEKSSEKGLRERAYGSSGELITSLPISIPLSTVQPNKLPVSIPLASVVLPSRAERARSTPSPVLQPRDPSSTLEKQIGANAHGAGSRSLALAPAGFSYAGSVAISGALAGSPASLTPGAEPATLDESSSSGSLFATVGSRSSTPQHPLLLAQPRNSLPASPAHQLSSSPRLGGAAQGPLPEASKGDLPSDSGFSDPESEAKRRIVFTITTGAGSAKQSPSSKHSPLTASARGDCVPSHGQDSRRRGRRKRASAGTPSLSAGVSPKRRALPSVAGLFTQPSGSPLNLNSMVSNINQPLEITAISSPETSLKSSPVPYQDHDQPPVLKKERPLSQTNGAHYSPLTSDEEPGSEDEPSSARIERKIATISLESKSPPKTLENGGGLAGRKPAPAGEPVNSSKWKSTFSPISDIGLAKSADSPLQASSALSQNSLFTFRPALEEPSADAKLAAHPRKGFPGSLSGADGLSPGTNPANGCTFGGGLAADLSLHSFSDGASLPHKGPEAAGLSSPLSFPSQRGKEGSDANPFLSKRQLDGLAGLKGEGSRGKEAGEGGLPLCGPTDKTPLLSGKAAKARDREVDLKNGHNLFISAAAVPPGSLLSGPGLAPAASSAGGAASSAQTHRSFLGPFPPGPQFALGPMSLQANLGSVAGSSVLQSLFSSVPAAAGLVHVSSAATRLTNSHAMGSFSGVAGGTVGGN af9_human Protein AF-9 ALL1-fused gene from chromosome 9 protein Myeloid/lymphoid or mixed-lineage leukemia translocated to chromosome 3 protein YEATS domain-containing protein 3 MLLT3 AF9 YEATS3 protein protein human Homo sapiens Human MASSCAVQVKLELGHRAQVRKKPTVEGFTHDWMVFVRGPEHSNIQHFVEKVVFHLHESFPRPKRVCKDPPYKVEESGYAGFILPIEVYFKNKEEPRKVRFDYDLFLHLEGHPPVNHLRCEKLTFNNPTEDFRRKLLKAGGDPNRSIHTSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSTSFSKPHKLMKEHKEKPSKDSREHKSAFKEPSRDHNKSSKESSKKPKENKPLKEEKIVPKMAFKEPKPMSKEPKPDSNLLTITSGQDKKAPSKRPPISDSEELSAKKRKKSSSEALFKSFSSAPPLILTCSADKKQIKDKSHVKMGKVKIESETSEKKKSTLPPFDDIVDPNDSDVEENISSKSDSEQPSPASSSSSSSSSFTPSQTRQQGPLRSIMKDLHSDDNEEESDEVEDNDNDSEMERPVNRGGSRSRRVSLSDGSDSESSSASSPLHHEPPPPLLKTNNNQILEVKSPIKQSKSDKQIKNGECDKAYLDELVELHRRLMTLRERHILQQIVNLIEETGHFHITNTTFDFDLCSLDKTTVRKLQSYLETSGTS enl_human Protein ENL MLLT1 ENL LTG19 YEATS1 YEATS domain-containing protein 1 protein protein human Homo sapiens Human MDNQCTVQVRLELGHRAQLRKKPTTEGFTHDWMVFVRGPEQCDIQHFVEKVVFWLHDSFPKPRRVCKEPPYKVEESGYAGFIMPIEVHFKNKEEPRKVCFTYDLFLNLEGNPPVNHLRCEKLTFNNPTTEFRYKLLRAGGVMVMPEGADTVSRPSPDYPMLPTIPLSAFSDPKKTKPSHGSKDANKESSKTSKPHKVTKEHRERPRKDSESKSSSKELEREQAKSSKDTSRKLGEGRLPKEEKAPPPKAAFKEPKMALKETKLESTSPKGGPPPPPPPPPRASSKRPATADSPKPSAKKQKKSSSKGSRSAPGTSPRTSSSSSFSDKKPAKDKSSTRGEKVKAESEPREAKKALEVEESNSEDEASFKSESAQSSPSNSSSSSDSSSDSDFEPSQNHSQGPLRSMVEDLQSEESDEDDSSSGEEAAGKTNPGRDSRLSFSDSESDNSADSSLPSREPPPPQKPPPPNSKVSGRRSPESCSKPEKILKKGTYDKAYTDELVELHRRLMALRERNVLQQIVNLIEETGHFNVTNTTFDFDLFSLDETTVRKLQSCLEAVAT aff4_human AF4/FMR2 family member 4 AFF4 AF5Q31 MCEF ALL1-fused gene from chromosome 5q31 protein Major CDK9 elongation factor-associated protein HSPC092 protein protein human Homo sapiens Human MNREDRNVLRMKERERRNQEIQQGEDAFPPSSPLFAEPYKVTSKEDKLSSRIQSMLGNYDEMKDFIGDRSIPKLVAIPKPTVPPSADEKSNPNFFEQRHGGSHQSSKWTPVGPAPSTSQSQKRSSGLQSGHSSQRTSAGSSSGTNSSGQRHDRESYNNSGSSSRKKGQHGSEHSKSRSSSPGKPQAVSSLNSSHSRSHGNDHHSKEHQRSKSPRDPDANWDSPSRVPFSSGQHSTQSFPPSLMSKSNSMLQKPTAYVRPMDGQESMEPKLSSEHYSSQSHGNSMTELKPSSKAHLTKLKIPSQPLDASASGDVSCVDEILKEMTHSWPPPLTAIHTPCKTEPSKFPFPTKESQQSNFGTGEQKRYNPSKTSNGHQSKSMLKDDLKLSSSEDSDGEQDCDKTMPRSTPGSNSEPSHHNSEGADNSRDDSSSHSGSESSSGSDSESESSSSDSEANEPSQSASPEPEPPPTNKWQLDNWLNKVNPHKVSPASSVDSNIPSSQGYKKEGREQGTGNSYTDTSGPKETSSATPGRDSKTIQKGSESGRGRQKSPAQSDSTTQRRTVGKKQPKKAEKAAAEEPRGGLKIESETPVDLASSMPSSRHKAATKGSRKPNIKKESKSSPRPTAEKKKYKSTSKSSQKSREIIETDTSSSDSDESESLPPSSQTPKYPESNRTPVKPSSVEEEDSFFRQRMFSPMEEKELLSPLSEPDDRYPLIVKIDLNLLTRIPGKPYKETEPPKGEKKNVPEKHTREAQKQASEKVSNKGKRKHKNEDDNRASESKKPKTEDKNSAGHKPSSNRESSKQSAAKEKDLLPSPAGPVPSKDPKTEHGSRKRTISQSSSLKSSSNSNKETSGSSKNSSSTSKQKKTEGKTSSSSKEVKEKAPSSSSNCPPSAPTLDSSKPRRTKLVFDDRNYSADHYLQEAKKLKHNADALSDRFEKAVYYLDAVVSFIECGNALEKNAQESKSPFPMYSETVDLIKYTMKLKNYLAPDATAADKRLTVLCLRCESLLYLRLFKLKKENALKYSKTLTEHLKNSYNNSQAPSPGLGSKAVGMPSPVSPKLSPGNSGNYSSGASSASASGSSVTIPQKIHQMAASYVQVTSNFLYATEIWDQAEQLSKEQKEFFAELDKVMGPLIFNASIMTDLVRYTRQGLHWLRQDAKLIS mll_enl_human_protein Fusion of human MLL (N-terminal region) with human ENL C-terminal residues 373-559 protein protein human Homo sapiens Human An amino acid sequence which is not represented in UniProt aff1_human AF4/FMR2 family member 1 AFF1 AF4 FEL MLLT2 PBM1 ALL1-fused gene from chromosome 4 protein Proto-oncogene AF4 Protein FEL protein protein human Homo sapiens Human MAAQSSLYNDDRNLLRIREKERRNQEAHQEKEAFPEKIPLFGEPYKTAKGDELSSRIQNMLGNYEEVKEFLSTKSHTHRLDASENRLGKPKYPLIPDKGSSIPSSSFHTSVHHQSIHTPASGPLSVGNISHNPKMAQPRTEPMPSLHAKSCGPPDSQHLTQDRLGQEGFGSSHHKKGDRRADGDHCASVTDSAPERELSPLISLPSPVPPLSPIHSNQQTLPRTQGSSKVHGSSNNSKGYCPAKSPKDLAVKVHDKETPQDSLVAPAQPPSQTFPPPSLPSKSVAMQQKPTAYVRPMDGQDQAPSESPELKPLPEDYRQQTFEKTDLKVPAKAKLTKLKMPSQSVEQTYSNEVHCVEEILKEMTHSWPPPLTAIHTPSTAEPSKFPFPTKDSQHVSSVTQNQKQYDTSSKTHSNSQQGTSSMLEDDLQLSDSEDSDSEQTPEKPPSSSAPPSAPQSLPEPVASAHSSSAESESTSDSDSSSDSESESSSSDSEENEPLETPAPEPEPPTTNKWQLDNWLTKVSQPAAPPEGPRSTEPPRRHPESKGSSDSATSQEHSESKDPPPKSSSKAPRAPPEAPHPGKRSCQKSPAQQEPPQRQTVGTKQPKKPVKASARAGSRTSLQGEREPGLLPYGSRDQTSKDKPKVKTKGRPRAAASNEPKPAVPPSSEKKKHKSSLPAPSKALSGPEPAKDNVEDRTPEHFALVPLTESQGPPHSGSGSRTSGCRQAVVVQEDSRKDRLPLPLRDTKLLSPLRDTPPPQSLMVKITLDLLSRIPQPPGKGSRQRKAEDKQPPAGKKHSSEKRSSDSSSKLAKKRKGEAERDCDNKKIRLEKEIKSQSSSSSSSHKESSKTKPSRPSSQSSKKEMLPPPPVSSSSQKPAKPALKRSRREADTCGQDPPKSASSTKSNHKDSSIPKQRRVEGKGSRSSSEHKGSSGDTANPFPVPSLPNGNSKPGKPQVKFDKQQADLHMREAKKMKQKAELMTDRVGKAFKYLEAVLSFIECGIATESESQSSKSAYSVYSETVDLIKFIMSLKSFSDATAPTQEKIFAVLCMRCQSILNMAMFRCKKDIAIKYSRTLNKHFESSSKVAQAPSPCIASTGTPSPLSPMPSPASSVGSQSSAGSVGSSGVAATISTPVTIQNMTSSYVTITSHVLTAFDLWEQAEALTRKNKEFFARLSTNVCTLALNSSLVDLVHYTRQGFQQLQELTKTP mll_aff4_fusion-1 Fusion of Human MLL (N -terminal region) and Human AFF4 ( C terminal region). protein protein human Homo sapiens Human An amino acid sequence which is not represented in UniProt mll_aff4_fusion Fusion of Human MLL (N -terminus) and Human AFF4 (518-1163) -C terminal region. protein protein human Homo sapiens Human An amino acid sequence which is not represented in UniProt ccnt1_human Cyclin-T1 CCNT1 protein protein human Homo sapiens Human MEGERKNNNKRWYFTREQLENSPSRRFGVDPDKELSYRQQAANLLQDMGQRLNVSQLTINTAIVYMHRFYMIQSFTQFPGNSVAPAALFLAAKVEEQPKKLEHVIKVAHTCLHPQESLPDTRSEAYLQQVQDLVILESIILQTLGFELTIDHPHTHVVKCTQLVRASKDLAQTSYFMATNSLHLTTFSLQYTPPVVACVCIHLACKWSNWEIPVSTDGKHWWEYVDATVTLELLDELTHEFLQILEKTPNRLKRIWNWRACEAAKKTKADDRGTDEKTSEQTILNMISQSSSDTTIAGLMSMSTSTTSAVPSLPVSEESSSNLTSVEMLPGKRWLSSQPSFKLEPTQGHRTSENLALTGVDHSLPQDGSNAFISQKQNSKSVPSAKVSLKEYRAKHAEELAAQKRQLENMEANVKSQYAYAAQNLLSHHDSHSSVILKMPIEGSENPERPFLEKADKTALKMRIPVAGGDKAASSKPEEIKMRIKVHAAADKHNSVEDSVTKSREHKEKHKTHPSNHHHHHNHHSHKHSHSQLPVGTGNKRPGDPKHSSQTSNLAHKTYSLSSSFSSSSSTRKRGPSEETGGAVFDHPAKIAKSTKSSSLNFSFPSLPTMGQMPGHSSDTSGLSFSQPSCKTRVPHSKLDKGPTGANGHNTTQTIDYQDTVNMLHSLLSAQGVQPTQPTAFEFVRPYSDYLNPRSGGISSRSGNTDKPRPPPLPSEPPPPLPPLPK men1_human Menin MEN1 SCG2 protein protein human Homo sapiens Human MGLKAAQKTLFPLRSIDDVVRLFAAELGREEPDLVLLSLVLGFVEHFLAVNRVIPTNVPELTFQPSPAPDPPGGLTYFPVADLSIIAALYARFTAQIRGAVDLSLYPREGGVSSRELVKKVSDVIWNSLSRSYFKDRAHIQSLFSFITGWSPVGTKLDSSGVAFAVVGACQALGLRDVHLALSEDHAWVVFGPNGEQTAEVTWHGKGNEDRRGQTVNAGVAERSWLYLKGSYMRCDRKMEVAFMVCAINPSIDLHTDSLELLQLQQKLLWLLYDLGHLERYPMALGNLADLEELEPTPGRPDPLTLYHKGIASAKTYYRDEHIYPYMYLAGYHCRNRNVREALQAWADTATVIQDYNYCREDEEIYKEFFEVANDVIPNLLKEAASLLEAGEERPGEQSQGTQSQGSALQDPECFAHLLRFYDGICKWEEGSPTPVLHVGWATFLVQSLGRFEGQVRQKVRIVSREAEAAEAEEPWGEEAREGRRRGPRRESKPEEPPPPKKPALDKGLGTGQGAVSGPPRKPPGTVAGTARGPEGGSTAQVPAPTASPPPEGPVLTFQSEKMKGMKELLVATKINSSAIKLQLTAQSQVQMKKQKVSTPSDYTLSFLKRQRKGL q03164-pro_0000390949 MLL cleavage product N320 KMT2A ALL-1 ALL1 CXXC-type zinc finger protein 7 CXXC7 HRX HTRX MLL MLL1 Myeloid/lymphoid or mixed-lineage leukemia Myeloid/lymphoid or mixed-lineage leukemia protein 1 TRX1 Trithorax-like protein Zinc finger protein HRX Cysteine methyltransferase KMT2A protein protein human Homo sapiens Human MAHSCRWRFPARPGTTGGGGGGGRRGLGGAPRQRVPALLLPPGPPVGGGGPGAPPSPPAVAAAAAAAGSSGAGVPGGAAAASAASSSSASSSSSSSSSASSGPALLRVGPGFDAALQVSAAIGTNLRRFRAVFGESGGGGGSGEDEQFLGFGSDEEVRVRSPTRSPSVKTSPRKPRGRPRSGSDRNSAILSDPSVFSPLNKSETKSGDKIKKKDSKSIEKKRGRPPTFPGVKIKITHGKDISELPKGNKEDSLKKIKRTPSATFQQATKIKKLRAGKLSPLKSKFKTGKLQIGRKGVQIVRRRGRPPSTERIKTPSGLLINSELEKPQKVRKDKEGTPPLTKEDKTVVRQSPRRIKPVRIIPSSKRTDATIAKQLLQRAKKGAQKKIEKEAAQLQGRKVKTQVKNIRQFIMPVVSAISSRIIKTPRRFIEDEDYDPPIKIARLESTPNSRFSAPSCGSSEKSSAASQHSSQMSSDSSRSSSPSVDTSTDSQASEEIQVLPEERSDTPEVHPPLPISQSPENESNDRRSRRYSVSERSFGSRTTKKLSTLQSAPQQQTSSSPPPPLLTPPPPLQPASSISDHTPWLMPPTIPLASPFLPASTAPMQGKRKSILREPTFRWTSLKHSRSEPQYFSSAKYAKEGLIRKPIFDNFRPPPLTPEDVGFASGFSASGTAASARLFSPLHSGTRFDMHKRSPLLRAPRFTPSEAHSRIFESVTLPSNRTSAGTSSSGVSNRKRKRKVFSPIRSEPRSPSHSMRTRSGRLSSSELSPLTPPSSVSSSLSISVSPLATSALNPTFTFPSHSLTQSGESAEKNQRPRKQTSAPAEPFSSSSPTPLFPWFTPGSQTERGRNKDKAPEELSKDRDADKSVEKDKSRERDREREKENKRESRKEKRKKGSEIQSSSALYPVGRVSKEKVVGEDVATSSSAKKATGRKKSSSHDSGTDITSVTLGDTTAVKTKILIKKGRGNLEKTNLDLGPTAPSLEKEKTLCLSTPSSSTVKHSTSSIGSMLAQADKLPMTDKRVASLLKKAKAQLCKIEKSKSLKQTDQPKAQGQESDSSETSVRGPRIKHVCRRAAVALGRKRAVFPDDMPTLSALPWEEREKILSSMGNDDKSSIAGSEDAEPLAPPIKPIKPVTRNKAPQEPPVKKGRRSRRCGQCPGCQVPEDCGVCTNCLDKPKFGGRNIKKQCCKMRKCQNLQWMPSKAYLQKQAKAVKKKEKKSKTSEKKDSKESSVVKNVVDSSQKPTPSAREDPAPKKSSSEPPPRKPVEEKSEEGNVSAPGPESKQATTPASRKSSKQVSQPALVIPPQPPTTGPPRKEVPKTTPSEPKKKQPPPPESGPEQSKQKKVAPRPSIPVKQKPKEKEKPPPVNKQENAGTLNILSTLSNGNSSKQKIPADGVHRIRVDFKEDCEAENVWEMGGLGILTSVPITPRVVCFLCASSGHVEFVYCQVCCEPFHKFCLEENERPLEDQLENWCCRRCKFCHVCGRQHQATKQLLECNKCRNSYHPECLGPNYPTKPTKKKKVWICTKCVRCKSCGSTTPGKGWDAQWSHDFSLCHDCAKLFAKGNFCPLCDKCYDDDDYESKMMQCGKCDRWVHSKCENLSDEMYEILSNLPESVAYTCVNCTERHPAEWRLALEKELQISLKQVLTALLNSRTTSHLLRYRQAAKPPDLNPETEESIPSRSSPEGPDPPVLTEVSKQDDQQPLDLEGVKRKMDQGNYTSVLEFSDDIVKIIQAAINSDGGQPEIKKANSMVKSFFIRQMERVFPWFSVKKSRFWEPNKVSSNSGMLPNAVLPPSLDHNYAQWQEREENSHTEQPPLMKKIIPAPKPKGPGEPDSPTPLHPPTPPILSTDRSREDSPELNPPPGIEDNRQCALCLTYGDDSANDAGRLLYIGQNEWTHVNCALWSAEVFEDDDGSLKNVHMAVIRGKQLRCEFCQKPGATVGCCLTSCTSNYHFMCSRAKNCVFLDDKKVYCQRHRDLIKGEVVPENGFEVFRRVFVDFEGISLRRKFLNGLEPENIHMMIGSMTIDCLGILNDLSDCEDKLFPIGYQCSRVYWSTTDARKRCVYTCKIVECRPPVVEPDINSTVEHDENRTIAHSPTSFTESSSKESQNTAEIISPPSPDRPPHSQTSGSCYYHVISKVPRIRTPSYSPTQRSPGCRPLPSAGSPTPTTHEIVTVGDPLLSSGLRSIGSRRHSTSSLSPQRSKLRIMSPMRTGNTYSRNNVSSVSTTGTATDLESSAKVVDHVLGPLNSSTSLGQNTSTSSNLQRTVVTVGNKNSHLDGSSSSEMKQSSASDLVSKSSSLKGEKTKVLSSKSSEGSAHNVAYPGIPKLAPQVHNTTSRELNVSKIGSFAEPSSVSFSSKEALSFPHLHLRGQRNDRDQHTDSTQSANSSPDEDTEVKTLKLSGMSNRSSIINEHMGSSSRDRRQKGKKSCKETFKEKHSSKSFLEPGQVTTGEEGNLKPEFMDEVLTPEYMGQRPCNNVSSDKIGDKGLSMPGVPKAPPMQVEGSAKELQAPRKRTVKVTLTPLKMENESQSKNALKESSPASPLQIESTSPTEPISASENPGDGPVAQPSPNNTSCQDSQSNNYQNLPVQDRNLMLPDGPKPQEDGSFKRRYPRRSARARSNMFFGLTPLYGVRSYGEEDIPFYSSSTGKKRGKRSAEGQVDGADDLSTSDEDDLYYYNFTRTVISSGGEERLASHNLFREEEQCDLPKISQLD Amino acids 1 � 2718. An N-terminal cleavage product of 320 kDa, identifier Q03164- PRO_0000390949. Short name is p320. 1 2718 cdk9_human Cyclin-dependent kinase 9 CDK9 CDC2L4 Cell division protein kinase 9 Serine/threonine-protein kinase PITALRE Cell division cycle 2-like protein kinase 4 C-2K TAK Tat-associated kinase complex catalytic subunit protein protein human Homo sapiens Human MAKQYDSVECPFCDEVSKYEKLAKIGQGTFGEVFKARHRKTGQKVALKKVLMENEKEGFPITALREIKILQLLKHENVVNLIEICRTKASPYNRCKGSIYLVFDFCEHDLAGLLSNVLVKFTLSEIKRVMQMLLNGLYYIHRNKILHRDMKAANVLITRDGVLKLADFGLARAFSLAKNSQPNRYTNRVVTLWYRPPELLLGERDYGPPIDLWGAGCIMAEMWTRSPIMQGNTEQHQLALISQLCGSITPEVWPNVDNYELYEKLELVKGQKRKVKDRLKAYVRDPYALDLIDKLLVLDPAQRIDSDDALNHDFFWSDPMPSDLKGMLSTHLTSMFEYLAPPRRKGSQITQQSTNQSRNPATTNQTEFERVF q03164-pro_0000390950 MLL cleavage product C180 KMT2A ALL-1 ALL1 CXXC-type zinc finger protein 7 CXXC7 HRX HTRX MLL MLL1 Myeloid/lymphoid or mixed-lineage leukemia Myeloid/lymphoid or mixed-lineage leukemia protein 1 TRX1 Trithorax-like protein Zinc finger protein HRX Cysteine methyltransferase KMT2A protein protein human Homo sapiens Human GVDDGTESDTSVTATTRKSSQIPKRNGKENGTENLKIDRPEDAGEKEHVTKSSVGHKNEPKMDNCHSVSRVKTQGQDSLEAQLSSLESSRRVHTSTPSDKNLLDTYNTELLKSDSDNNNSDDCGNILPSDIMDFVLKNTPSMQALGESPESSSSELLNLGEGLGLDSNREKDMGLFEVFSQQLPTTEPVDSSVSSSISAEEQFELPLELPSDLSVLTTRSPTVPSQNPSRLAVISDSGEKRVTITEKSVASSESDPALLSPGVDPTPEGHMTPDHFIQGHMDADHISSPPCGSVEQGHGNNQDLTRNSSTPGLQVPVSPTVPIQNQKYVPNSTDSPGPSQISNAAVQTTPPHLKPATEKLIVVNQNMQPLYVLQTLPNGVTQKIQLTSSVSSTPSVMETNTSVLGPMGGGLTLTTGLNPSLPTSQSLFPSASKGLLPMSHHQHLHSFPAATQSSFPPNISNPPSGLLIGVQPPPDPQLLVSESSQRTDLSTTVATPSSGLKKRPISRLQTRKNKKLAPSSTPSNIAPSDVVSNMTLINFTPSQLPNHPSLLDLGSLNTSSHRTVPNIIKRSKSSIMYFEPAPLLPQSVGGTAATAAGTSTISQDTSHLTSGSVSGLASSSSVLNVVSMQTTTTPTSSASVPGHVTLTNPRLLGTPDIGSISNLLIKASQQSLGIQDQPVALPPSSGMFPQLGTSQTPSTAAITAASSICVLPSTQTTGITAASPSGEADEHYQLQHVNQLLASKTGIHSSQRDLDSASGPQVSNFTQTVDAPNSMGLEQNKALSSAVQASPTSPGGSPSSPSSGQRSASPSVPGPTKPKPKTKRFQLPLDKGNGKKHKVSHLRTSSSEAHIPDQETTSLTSGTGTPGAEAEQQDTASVEQSSQKECGQPAGQVAVLPEVQVTQNPANEQESAEPKTVEEEESNFSSPLMLWLQQEQKRKESITEKKPKKGLVFEISSDDGFQICAESIEDAWKSLTDKVQEARSNARLKQLSFAGVNGLRMLGILHDAVVFLIEQLSGAKHCRNYKFRFHKPEEANEPPLNPHGSARAEVHLRKSAFDMFNFLASKHRQPPEYNPNDEEEEEVQLKSARRATSMDLPMPMRFRHLKKTSKEAVGVYRSPIHGRGLFCKRNIDAGEMVIEYAGNVIRSIQTDKREKYYDSKGIGCYMFRIDDSEVVDATMHGNAARFINHSCEPNCYSRVINIDGQKHIVIFAMRKIYRGEELTYDYKFPIEDASNKLPCNCGAKKCRKFLN Amino acids 2719 � 3969. An C-terminal cleavage product of 180 kDa, identifier Q03164-PRO_0000390950. Short name is p180. 2719 3969 q2q440_human protein protein human Homo sapiens Human FLCASSGHVEDANKESSKTS q8td98_human protein protein human Homo sapiens Human TTGPPRKEVPKTTPSEPKKKQPPPPESGPEQSKQKKVAPRPSIPVKQKPKEKEKPPPVNKQENAGTLNILSTLSNGNSSKQKIPADGVHRIRVDFKEDCEAENVWEMGGLGILTSVPITPRVVCFLCASSGHVEQTYSNEVHCVEEILKEMTHSWPPPLAAIHTPSTAEPSKFPFPTKDSQHVSSVTQNQKQYDTSSKTHSNSQQGTSSMLEDDLQLSDSEDSDSEQTPEKPPSHLHLQVLHSPFQNQWHQHIPAVQSQKAPVTQTVPQTQRARAVQVTASSSDSEENEPLETPAPEPEPPTTNKWQLDNWLTKVSQPAAPPEGPRSTEPPRRHPESKGSSDSATSQEHSESKDPPPKSSSKAPRAPPEAPHPGKRSCQKSPAQQEPPQRQTVGTKQPKKPVKASARAGSRTSLQGEREPGLLPYGSRDQTSKDKPKVKTKGRPRAAASNEPKPAVPPSSEKKKHKSSLPAPSKALSGPEPAKDNVEDRTPEHFALVPLTESQGPPHSGSGSRTSGCRQAVVVQEDSRKDRLPLPLRDTKLLSPLRDTPPPQSLMVKITLDLLSRIPQPPGKGSRQRKAEDKQPPAGKKHSSEKRSSDSSSKLAKKRKGEAERDCDNKKIRLEK q1wd23_human MLL/AF6 fusion protein protein human Homo sapiens Human RPSIPVKQKPKEKEKPPPVNKQENAGTLNILSTLSNGNSSKQKIPADGVHRIRVDFKDLEFHGVMRFYFQDKAAGNFATKCIRVSSTATTQDVIETLAEKFRPDMRMLSSPKYSLYEVHVSGERRL af9-dot1l 886646 n/a 886655 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 886656 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P42568:p.Leu558Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false gal4bd GAL4 DNA binding domain fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 4D. The AHD of ENL also mediated association with DOT1L. This (AHD) region is evolutionally conserved in AF9. enl-dot1l 886646 n/a 886655 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 886657 unspecified role unspecified role bait bait Q03111:p.Leu550Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false gal4bd GAL4 DNA binding domain fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c-terminal 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 4D. The AHD of ENL also mediated association with DOT1L. dot1l-enl 886646 n/a 886657 unspecified role unspecified role prey prey xpress-tag Asp-Leu-Tyr-Asp-Asp-Asp-Asp-Lys Tag fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 886655 unspecified role unspecified role bait bait 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 Fig 5A. ENL coprecipitated both AFF4/AF5q31 and DOT1L, but the latter two did not pull down each other, indicating that the three proteins do not form a trimeric complex. enl-dot1l-1 886646 n/a 886658 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 n/a 886655 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 886657 unspecified role unspecified role bait bait xpress-tag Asp-Leu-Tyr-Asp-Asp-Asp-Asp-Lys Tag fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false Fig 5A. ENL coprecipitated both AFF4/AF5q31 and DOT1L, but the latter two did not pull down each other, indicating that the three proteins do not form a trimeric complex. aff4-enl 886646 n/a 886657 unspecified role unspecified role prey prey xpress-tag Asp-Leu-Tyr-Asp-Asp-Asp-Asp-Lys Tag fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 886658 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 5A. ENL coprecipitated both AFF4/AF5q31 and DOT1L, but the latter two did not pull down each other, indicating that the three proteins do not form a trimeric complex. mll-enl-aff1 886646 n/a 886659 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 886655 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 886660 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 association association false false false Fig 5C. FLAG-tagged MLL fusion proteins were coexpressed with AFF1 or DOT1L. mll-aff4-aff1 886646 n/a 886661 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 886660 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 physical association physical association false false false Fig 5C. Artificial MLL fusion protein (with AFF4) that does associate with AFF1. mll-aff4-34region 886646 n/a 886662 unspecified role unspecified role prey prey xpress-tag Asp-Leu-Tyr-Asp-Asp-Asp-Asp-Lys Tag fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 886663 unspecified role unspecified role prey prey n/a 886660 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false Fig S3A. Preferential hetero-dimerization was also observed in co-IP experiments of endogenous or transfected MLL-AFF4(AF5q31). aff4-ct1 886646 n/a 886658 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 886663 unspecified role unspecified role prey prey physical association physical association false false false Fig S3A. Preferential hetero-dimerization was also observed in co-IP experiments of endogenous or transfected MLL-AFF4(AF5q31). enl-dot1l-2 886646 n/a 886655 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 886657 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false gal4bd GAL4 DNA binding domain fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig S4. AFF4 co-precipitates ENL but not DOT1L under conditions where ENL pulls down DOT1L. aff4-enl-1 886646 n/a 886657 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false gal4bd GAL4 DNA binding domain fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 886658 unspecified role unspecified role bait bait subregion_3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false gal4bd GAL4 DNA binding domain fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false Fig S4. AFF4 co-precipitates ENL but not DOT1L under conditions where ENL pulls down DOT1L. men1-mlln 886647 n/a 886664 unspecified role unspecified role prey prey n/a 886665 unspecified role unspecified role bait bait physical association physical association false false false Fig 1C. MLL did not pull down AEP components in K562 cells while coprecipitating menin, a component of the MLL complex. af4-3 886647 n/a 886658 unspecified role unspecified role prey prey n/a 886657 unspecified role unspecified role prey prey n/a 886666 unspecified role unspecified role prey prey n/a 886663 unspecified role unspecified role prey prey n/a 886660 unspecified role unspecified role bait bait association association false false false Fig 1C. Reciprocal immunoprecipitation further confirmed that all five proteins compose an endogenous complex consistent with previous observations. af4-4 886647 n/a 886666 unspecified role unspecified role prey prey n/a 886663 unspecified role unspecified role prey prey n/a 886658 unspecified role unspecified role bait bait n/a 886657 unspecified role unspecified role prey prey association association false false false Fig 1C. Reciprocal immunoprecipitation further confirmed that all five proteins compose an endogenous complex consistent with previous observations. af4-5 886647 n/a 886657 unspecified role unspecified role bait bait n/a 886658 unspecified role unspecified role prey prey n/a 886663 unspecified role unspecified role prey prey n/a 886666 unspecified role unspecified role prey prey association association false false false Fig 1C. Reciprocal immunoprecipitation further confirmed that all five proteins compose an endogenous complex consistent with previous observations. mlln-mllc 886647 n/a 886667 unspecified role unspecified role prey prey n/a 886665 unspecified role unspecified role bait bait physical association physical association false false false Fig 1C. Reciprocal immunoprecipitation further confirmed that all five proteins compose an endogenous complex consistent with previous observations. af4-7 886647 n/a 886666 unspecified role unspecified role bait bait n/a 886660 unspecified role unspecified role prey prey n/a 886663 unspecified role unspecified role prey prey n/a 886657 unspecified role unspecified role prey prey n/a 886658 unspecified role unspecified role prey prey association association false false false Fig 1C. Reciprocal immunoprecipitation further confirmed that all five proteins compose an endogenous complex consistent with previous observations. aff1-ptefb 886648 n/a 886666 unspecified role unspecified role prey prey n/a 886663 unspecified role unspecified role prey prey n/a 886660 unspecified role unspecified role bait bait gal4bd GAL4 DNA binding domain fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false subregion_1 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false association association false false false Fig 3B. P-TEFb interacts with AFF1/AF4 and AFF4/AF5q31 via subregion 1, which contains the N-terminal homology domain (NHD). aff4-ptefb 886648 n/a 886666 unspecified role unspecified role prey prey n/a 886658 unspecified role unspecified role bait bait subregion_1 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false gal4bd GAL4 DNA binding domain fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 886663 unspecified role unspecified role prey prey association association false false false Fig 3B. P-TEFb interacts with AFF1/AF4 and AFF4/AF5q31 via subregion 1, which contains the N-terminal homology domain (NHD). enl-aff4 886648 n/a 886657 unspecified role unspecified role bait bait Q03111:p.Leu550Glu mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false gal4bd GAL4 DNA binding domain fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c-terminal required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false n/a 886658 unspecified role unspecified role prey prey physical association physical association false false false Fig 4B. C-terminal ENL residues (494�559) are required for the interaction with (endogenous) AFF4/AF5q31. af9-aff4 886648 n/a 886658 unspecified role unspecified role prey prey n/a 886656 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P42568:p.Leu558Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false gal4bd GAL4 DNA binding domain fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 4B. C-terminal ENL residues (494�559) are required for the interaction with (endogenous) AFF4/AF5q31. This (AHD) region is evolutionally conserved in AF9. af4-8 886649 n/a 886665 unspecified role unspecified role bait bait n/a 886668 unspecified role unspecified role prey prey n/a 886658 unspecified role unspecified role prey prey n/a 886663 unspecified role unspecified role prey prey n/a 886666 unspecified role unspecified role prey prey n/a 886660 unspecified role unspecified role prey prey association association false false false Fig 1E. Reciprocal IP using human leukemia cell lines that express MLL-ENL, MLL-AF4, or MLL-AF5q31 showed that the respective fusion proteins form similar AEP-like complexes. af4-9 886649 n/a 886657 unspecified role unspecified role prey prey n/a 886660 unspecified role unspecified role bait bait n/a 886663 unspecified role unspecified role prey prey n/a 886658 unspecified role unspecified role prey prey n/a 886666 unspecified role unspecified role prey prey n/a 886668 unspecified role unspecified role prey prey association association false false false Fig 1E. Reciprocal IP using human leukemia cell lines that express MLL-ENL, MLL-AF4, or MLL-AF5q31 showed that the respective fusion proteins form similar AEP-like complexes. af4-10 886650 n/a 886669 unspecified role unspecified role prey prey n/a 886665 unspecified role unspecified role bait bait n/a 886657 unspecified role unspecified role prey prey n/a 886666 unspecified role unspecified role prey prey n/a 886660 unspecified role unspecified role prey prey n/a 886663 unspecified role unspecified role prey prey association association false false false Fig 1E. Reciprocal IP using human leukemia cell lines that express MLL-ENL, MLL-AF4, or MLL-AF5q31 showed that the respective fusion proteins form similar AEP-like complexes. af4-11 886650 n/a 886669 unspecified role unspecified role prey prey n/a 886658 unspecified role unspecified role prey prey n/a 886660 unspecified role unspecified role bait bait n/a 886663 unspecified role unspecified role prey prey n/a 886666 unspecified role unspecified role prey prey n/a 886657 unspecified role unspecified role prey prey association association false false false Fig 1E. Reciprocal IP using human leukemia cell lines that express MLL-ENL, MLL-AF4, or MLL-AF5q31 showed that the respective fusion proteins form similar AEP-like complexes. mlln-mlaf6 886651 n/a 886670 unspecified role unspecified role prey prey n/a 886665 unspecified role unspecified role bait bait physical association physical association false false false Fig 1E. MLL-AF6, an MLL fusion with a cytoplasmic protein that was not copurified with AF4 or AF5q31, did not coprecipitate any of the AEP components in ML-2 cells. ML-2 cells have no normal MLL gene. af4-13 886651 n/a 886658 unspecified role unspecified role prey prey n/a 886657 unspecified role unspecified role prey prey n/a 886666 unspecified role unspecified role prey prey n/a 886660 unspecified role unspecified role bait bait n/a 886663 unspecified role unspecified role prey prey association association false false false Fig 1E. MLL-AF6, an MLL fusion with a cytoplasmic protein that was not copurified with AF4 or AF5q31, did not coprecipitate any of the AEP components in ML-2 cells. ML-2 cells have no normal MLL gene. aff1-ptefb-1 886652 n/a 886663 unspecified role unspecified role prey prey n/a 886666 unspecified role unspecified role prey prey n/a 886660 unspecified role unspecified role bait bait n/a 886658 unspecified role unspecified role prey prey n/a 886657 unspecified role unspecified role prey prey association association false false false Fig S1B. Reciprocal IP using human leukemia cell lines that express MLL-ENL, MLL-AFF1(AF4), or MLL-AFF1(AF5q31) showed that the respective fusion proteins form similar AEP-like complexes. mll-ptefb 886652 n/a 886665 unspecified role unspecified role bait bait n/a 886658 unspecified role unspecified role prey prey n/a 886657 unspecified role unspecified role prey prey n/a 886663 unspecified role unspecified role prey prey n/a 886666 unspecified role unspecified role prey prey association association false false false Fig S1B. Reciprocal IP using human leukemia cell lines that express MLL-ENL, MLL-AFF1(AF4), or MLL-AFF1(AF5q31) showed that the respective fusion proteins form similar AEP-like complexes. af4-aff1-1 886653 n/a 886666 unspecified role unspecified role prey prey n/a 886663 unspecified role unspecified role prey prey n/a 886657 unspecified role unspecified role prey prey n/a 886660 unspecified role unspecified role bait bait n/a 886658 unspecified role unspecified role prey prey association association false false false Fig 1B. In the identification of AF4-associated proteins in vivo, Mass spectrometry identified AFF4/AF5q31, ENL, CDK9, and cyclin T1 in the purified materials. af4-2 886654 n/a 886658 unspecified role unspecified role bait bait n/a 886657 unspecified role unspecified role prey prey n/a 886660 unspecified role unspecified role prey prey n/a 886663 unspecified role unspecified role prey prey n/a 886666 unspecified role unspecified role prey prey association association false false false Fig S1A. AFF1/AF4, AFF4/AF5q31, and ENL associate in an endogenous higher-order complex.