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/ liu-2006b-1 The tetramer structure of the Nervy homology two domain, NHR2, is critical for AML1/ETO's activity. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Liu Y., Cheney MD., Gaudet JJ., Chruszcz M., Lukasik SM., Sugiyama D., Lary J., Cole J., Dauter Z., Minor W., Speck NA., Bushweller JH. Cancer - Interactions investigated in the context of cancer nancy.speck@dartmouth.edu Cancer Cell (1535-6108) 2006 Only protein-protein interactions 3 imex curation liu-2006b-2 The tetramer structure of the Nervy homology two domain, NHR2, is critical for AML1/ETO's activity. in vitro In vitro cosedimentation cosedimentation predetermined predetermined participant predetermined Liu Y., Cheney MD., Gaudet JJ., Chruszcz M., Lukasik SM., Sugiyama D., Lary J., Cole J., Dauter Z., Minor W., Speck NA., Bushweller JH. Cancer - Interactions investigated in the context of cancer Absorbance data 280 nm (NHR2, m4), 230 nm (m7, m2) were collected at a rotor speed of 50, 000 rpm at 20 C in aluminum-filled double sector cells at a scan interval of approximately 260 s. nancy.speck@dartmouth.edu Cancer Cell (1535-6108) 2006 Only protein-protein interactions 3 imex curation liu-2006b-3 The tetramer structure of the Nervy homology two domain, NHR2, is critical for AML1/ETO's activity. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Liu Y., Cheney MD., Gaudet JJ., Chruszcz M., Lukasik SM., Sugiyama D., Lary J., Cole J., Dauter Z., Minor W., Speck NA., Bushweller JH. Cancer - Interactions investigated in the context of cancer nancy.speck@dartmouth.edu Cancer Cell (1535-6108) 2006 Only protein-protein interactions 3 imex curation mtg8_human Protein CBFA2T1 Protein MTG8 Protein ETO Eight twenty one protein Cyclin-D-related protein Zinc finger MYND domain-containing protein 2 RUNX1T1 AML1T1 CBFA2T1 CDR ETO MTG8 ZMYND2 protein protein human Homo sapiens Human MISVKRNTWRALSLVIGDCRKKGNFEYCQDRTEKHSTMPDSPVDVKTQSRLTPPTMPPPPTTQGAPRTSSFTPTTLTNGTSHSPTALNGAPSPPNGFSNGPSSSSSSSLANQQLPPACGARQLSKLKRFLTTLQQFGNDISPEIGERVRTLVLGLVNSTLTIEEFHSKLQEATNFPLRPFVIPFLKANLPLLQRELLHCARLAKQNPAQYLAQHEQLLLDASTTSPVDSSELLLDVNENGKRRTPDRTKENGFDREPLHSEHPSKRPCTISPGQRYSPNNGLSYQPNGLPHPTPPPPQHYRLDDMAIAHHYRDSYRHPSHRDLRDRNRPMGLHGTRQEEMIDHRLTDREWAEEWKHLDHLLNCIMDMVEKTRRSLTVLRRCQEADREELNYWIRRYSDAEDLKKGGGSSSSHSRQQSPVNPDPVALDAHREFLHRPASGYVPEEIWKKAEEAVNEVKRQAMTELQKAVSEAERKAHDMITTERAKMERTVAEAKRQAAEDALAVINQQEDSSESCWNCGRKASETCSGCNTARYCGSFCQHKDWEKHHHICGQTLQAQQQGDTPAVSSSVTPNSGAGSPMDTPPAATPRSTTPGTPSTIETTPR aml1_eto_chimera AML1-ETO chimera, human RUNX1(1-177)-N-MTG8(31-604) protein protein human Homo sapiens Human MRIPVDASTSRRFTPPSTALSPGKMSEALPLGAPDAGAALAGKLRSGDRSMVEVLADHPGELVRTDSPNFLCSVLPTHWRCNKTLPIAFKVVALGDVPDGTLVTVMAGNDENYSAELRNATAAMKNQVARFNDLRFVGRSGRGKSFTLTITVFTNPPQVATYHRAIKITVDGPREPRNRTEKHSTMPDSPVDVKTQSRLTPPTMPPPPTTQGAPRTSSFTPTTLTNGTSHSPTALNGAPSPPNGFSNGPSSSSSSSLANQQLPPACGARQLSKLKRFLTTLQQFGNDISPEIGERVRTLVLGLVNSTLTIEEFHSKLQEATNFPLRPFVIPFLKANLPLLQRELLHCARLAKQNPAQYLAQHEQLLLDASTTSPVDSSELLLDVNENGKRRTPDRTKENGFDREPLHSEHPSKRPCTISPGQRYSPNNGLSYQPNGLPHPTPPPPQHYRLDDMAIAHHYRDSYRHPSHRDLRDRNRPMGLHGTRQEEMIDHRLTDREWAEEWKHLDHLLNCIMDMVEKTRRSLTVLRRCQEADREELNYWIRRYSDAEDLKKGGGSSSSHSRQQSPVNPDPVALDAHREFLHRPASGYVPEEIWKKAEEAVNEVKRQAMTELQKAVSEAERKAHDMITTERAKMERTVAEAKRQAAEDALAVINQQEDSSESCWNCGRKASETCSGCNTARYCGSFCQHKDWEKHHHICGQTLQAQQQGDTPAVSSSVTPNSGAGSPMDTPPAATPRSTTPGTPSTIETTPR runx11 509086 N/A 509089 unspecified role unspecified role neutral component neutral component region SeMet substituted se(s)met L-selenomethionine Delta:S(-1)Se(1) Se(S)Met Selenium replaces sulphur semetm undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region NHR2 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 4.0 direct interaction direct interaction false false false Note the pdb entry refers to the Uniprot entry Q06455 (resides 335-403). However, the authors refer to the numbering for AML1_ETO (480-551). The protein crystallised has an extra ala-met-ala at the N-terminus. 1, Table 1 2.0 A R(%) 21.6; Rfree(%) 27.0 runx11-1 509087 N/A 509090 unspecified role unspecified role neutral component neutral component region NHR2 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false aml1_eto_chimera:p.[Leu493Arg;Leu505Glu;Leu508Glu;Leu509Arg;Leu523Arg;Leu526Glu;Leu537Arg] m7 mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 4.0 direct interaction direct interaction false false false The Uniprot entry Q06455 (resides 335-403)contains the NHR2 domain as does the chimeric protein AML1_ETO. 3 C, Table 2 The m4 (leu345gln-leu375glu-leu378lys-leu389gln) mutant was tetrameric at 20�C by analytical ultracentrifugation (Table 2), but CD spectroscopy revealed that it was only 41% helical, suggesting that the m4 tetramer is significantly destabilized. runx1-human-protein 509088 n/a 509090 unspecified role unspecified role neutral component neutral component aml1_eto_chimera:p.[Leu493Glu;Leu505Arg;Leu508Arg;Leu509Glu;Leu523Glu;Leu526Arg;Leu537Arg] m7 mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false aml1_eto_chimera:p.[Leu493Gln;Leu523Glu;Leu526Lys;Leu537Gln] m4 mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region deletion of the NHR2 domain required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false in vitro In vitro direct interaction direct interaction false false false 3 A, E It is not clear whether the deletion of the NHR2 domain completely abolishes oligomerisation of AML1_ETO. The elution volume of the m4 mutant is in between that for AML1_ETO and the m7 mutant.