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/ wang-2016-50 in vitro In vitro radiolabeled methyl methyltransferase radiometric assay radiolabeled methyl predetermined predetermined participant predetermined Wang P., Doxtader KA., Nam Y. Cancer - Interactions investigated in the context of cancer yunsun.nam@utsouthwestern.edu Molecular cell 2016 Only protein-protein interactions imex curation imex curation wang-2016-49 in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Wang P., Doxtader KA., Nam Y. Cancer - Interactions investigated in the context of cancer yunsun.nam@utsouthwestern.edu Molecular cell 2016 Only protein-protein interactions imex curation imex curation wang-2016-48 in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Wang P., Doxtader KA., Nam Y. Cancer - Interactions investigated in the context of cancer SEC-MALS -Size Exclusion Chromatography with Multi-Angle Light Scattering or SEC followed by SDS-PAGE and protein visualisation with Stain-Free dye (Biorad). yunsun.nam@utsouthwestern.edu Molecular cell 2016 Only protein-protein interactions imex curation imex curation “ggacu” “GGACU” rna ribonucleic acid RNA rna synthetic construct synthetic construct UACACUCGAUCUGGACUAAAGCUGCUC met14_human N6-adenosine-methyltransferase non-catalytic subunit Methyltransferase-like protein 14 METTL14 KIAA1627 protein protein human Homo sapiens Human MDSRLQEIRERQKLRRQLLAQQLGAESADSIGAVLNSKDEQREIAETRETCRASYDTSAPNAKRKYLDEGETDEDKMEEYKDELEMQQDEENLPYEEEIYKDSSTFLKGTQSLNPHNDYCQHFVDTGHRPQNFIRDVGLADRFEEYPKLRELIRLKDELIAKSNTPPMYLQADIEAFDIRELTPKFDVILLEPPLEEYYRETGITANEKCWTWDDIMKLEIDEIAAPRSFIFLWCGSGEGLDLGRVCLRKWGYRRCEDICWIKTNKNNPGKTKTLDPKAVFQRTKEHCLMGIKGTVKRSTDGDFIHANVDIDLIITEEPEIGNIEKPVEIFHIIEHFCLGRRRLHLFGRDSTIRPGWLTVGPTLTNSNYNAETYASYFSAPNSYLTGCTEEIERLRPKSPPPKSKSDRGGGAPRGGGRGGTSAGRGRERNRSNFRGERGGFRGGRGGAHRGGFPPR mta70_human N6-adenosine-methyltransferase catalytic subunit N6-adenosine-methyltransferase 70 kDa subunit METTL3 MTA70 Methyltransferase-like protein 3 protein protein human Homo sapiens Human MSDTWSSIQAHKKQLDSLRERLQRRRKQDSGHLDLRNPEAALSPTFRSDSPVPTAPTSGGPKPSTASAVPELATDPELEKKLLHHLSDLALTLPTDAVSICLAISTPDAPATQDGVESLLQKFAAQELIEVKRGLLQDDAHPTLVTYADHSKLSAMMGAVAEKKGPGEVAGTVTGQKRRAEQDSTTVAAFASSLVSGLNSSASEPAKEPAKKSRKHAASDVDLEIESLLNQQSTKEQQSKKVSQEILELLNTTTAKEQSIVEKFRSRGRAQVQEFCDYGTKEECMKASDADRPCRKLHFRRIINKHTDESLGDCSFLNTCFHMDTCKYVHYEIDACMDSEAPGSKDHTPSQELALTQSVGGDSSADRLFPPQWICCDIRYLDVSILGKFAVVMADPPWDIHMELPYGTLTDDEMRRLNIPVLQDDGFLFLWVTGRAMELGRECLNLWGYERVDEIIWVKTNQLQRIIRTGRTGHWLNHGKEHCLVGVKGNPQGFNQGLDCDVIVAEVRSTSHKPDEIYGMIERLSPGTRKIELFGRPHNVQPNWITLGNQLDGIHLLDPDVVARFKQRYPDGIISKPKNL s-adenosyl-l-homocysteine S-adenosyl-l-homocysteine small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) s-adenosyl-l-methionine S adenosyl Methionine small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) mettl14-mettl3-18 3326026 N/A 3326029 enzyme target enzyme target substrate substrate neutral component neutral component purified purified c16u mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false synthetic construct synthetic construct N/A 3326030 unspecified role unspecified role neutral component neutral component purified purified whole_molecule 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) N/A 3326031 enzyme enzyme neutral component neutral component purified purified Q86U44:p.Asp395Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false Q86U44:p.Trp475Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false Q86U44:p.Cys294Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false Q86U44:p.Tyr406Cys mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q86U44:p.Asn477Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false Q86U44:p.Cys326Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 3326044 3326038 methylation methylation reaction methylation false false false Figure 2D, 4H, 5A/D mettl14-mettl3-19 3326026 N/A 3326029 enzyme target enzyme target substrate substrate neutral component neutral component purified purified synthetic construct synthetic construct N/A 3326030 unspecified role unspecified role neutral component neutral component purified purified Q9HCE5:p.Asp312Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false Q9HCE5:p.Glu192Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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. N/A 3326031 enzyme enzyme neutral component neutral component purified purified whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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. 3326054 3326052 methylation methylation reaction methylation false false false Figure 2E, 4G/I Mettl3 is the catalytically active subunit, its activity is negligible without forming stoichiometric complexes with Mettl14. mettl14-mettl3-20 3326026 N/A 3326029 enzyme target enzyme target substrate substrate neutral component neutral component purified purified c16u mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false synthetic construct synthetic construct N/A 3326030 unspecified role unspecified role neutral component neutral component purified purified Q9HCE5:p.Asp312Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false Q9HCE5:p.Glu192Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false whole_molecule 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) N/A 3326031 enzyme enzyme neutral component neutral component purified purified whole_molecule 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) 3326063 3326061 methylation methylation reaction methylation false false false Figure 2D-2, 4H, 5A mettl14-mettl3-21 3326026 N/A 3326029 enzyme target enzyme target substrate substrate neutral component neutral component purified purified c16u mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false synthetic construct synthetic construct N/A 3326030 unspecified role unspecified role neutral component neutral component purified purified region_including_methyltransferase_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) N/A 3326031 enzyme enzyme neutral component neutral component purified purified whole_molecule 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) 3326070 3326068 methylation methylation reaction methylation false false false Figure 4H mettl14-mettl3-22 3326026 N/A 3326029 enzyme target enzyme target substrate substrate neutral component neutral component purified purified synthetic construct synthetic construct N/A 3326030 unspecified role unspecified role neutral component neutral component purified purified whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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. N/A 3326031 enzyme enzyme neutral component neutral component purified purified Q86U44:p.Tyr406Cys mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false Q86U44:p.Asp395Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q86U44:p.Trp475Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false Q86U44:p.Cys294Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false Q86U44:p.Cys326Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false Q86U44:p.Asn477Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false 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. 3326078 3326074 methylation methylation reaction methylation false false false Figure 2E-2, 4G/I,5B/E Mettl3 is the catalytically active subunit, its activity is negligible without forming stoichiometric complexes with Mettl14. mettl14-mettl3-23 3326026 N/A 3326029 enzyme target enzyme target substrate substrate neutral component neutral component purified purified synthetic construct synthetic construct N/A 3326030 unspecified role unspecified role neutral component neutral component purified purified region_including_methyltransferase_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) N/A 3326031 enzyme enzyme neutral component neutral component purified purified (ccch)x2_motifs required to bind necessary binding region required to bind range range range range false (ccch)x2_motifs+methyltransferase 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) 3326089 3326086 methylation methylation reaction methylation false false false Figure 5A (Supp fig 6A) mettl14-mettl3-24 3326026 N/A 3326029 enzyme target enzyme target substrate substrate neutral component neutral component purified purified c16u mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false synthetic construct synthetic construct N/A 3326030 unspecified role unspecified role neutral component neutral component purified purified Q9HCE5:p.Arg298Pro mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false whole_molecule 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) N/A 3326031 enzyme enzyme neutral component neutral component purified purified whole_molecule 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) 3326097 3326095 methylation methylation reaction methylation false false false Figure 5D, (Supp 3A) mettl14-mettl3-25 3326026 N/A 3326029 enzyme target enzyme target substrate substrate neutral component neutral component purified purified c16u mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false synthetic construct synthetic construct N/A 3326030 unspecified role unspecified role neutral component neutral component purified purified Q9HCE5:p.Arg298Pro mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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. N/A 3326031 enzyme enzyme neutral component neutral component purified purified whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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. 3326105 3326103 methylation methylation reaction methylation false false false Figure 5E, Supp 3A mettl14-mettl3-26 3326026 N/A 3326029 enzyme target enzyme target substrate substrate neutral component neutral component purified purified synthetic construct synthetic construct N/A 3326030 unspecified role unspecified role neutral component neutral component purified purified whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. N/A 3326031 enzyme enzyme neutral component neutral component purified purified whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 3326111 3326109 methylation methylation reaction methylation false false false Supp 3A/C mettl14-mettl3-15 3326027 N/A 3326032 cofactor cofactor coenzyme neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 N/A 3326030 unspecified role unspecified role neutral component neutral component purified purified region_including_methyltransferase_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: 1.0 N/A 3326031 unspecified role unspecified role neutral component neutral component purified purified methyltransferase_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: 1.0 direct interaction direct interaction false false false Table 1 1.65A R Work= 0.16 R Free= 0.179 mettl14-mettl3-16 3326027 N/A 3326033 cofactor cofactor coenzyme neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 N/A 3326030 unspecified role unspecified role neutral component neutral component purified purified region_including_methyltransferase_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: 1.0 N/A 3326031 unspecified role unspecified role neutral component neutral component purified purified methyltransferase_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: 1.0 direct interaction direct interaction false false false Table 1i 1.7A R Work= 0.163 R Free= 0.187 mettl14-mettl3-17 3326027 N/A 3326030 unspecified role unspecified role neutral component neutral component purified purified region_including_methyltransferase_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: 1.0 N/A 3326031 unspecified role unspecified role neutral component neutral component purified purified methyltransferase_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: 1.0 direct interaction direct interaction false false false Table 1ii 1.65A R Work= 0.16 R Free= 0.184 mettl14-mettl3-14 3326028 N/A 3326031 unspecified role unspecified role neutral component neutral component purified purified methyltransferase_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: 1.0 N/A 3326030 unspecified role unspecified role neutral component neutral component purified purified region_including_methyltransferase_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: 1.0 direct interaction direct interaction false false false Figure 1B/C, Supp. figure 1A/C