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/ peng-2017-1 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 Peng M., Yin N., Li MO. Results from Amino acid binding assay have not been included as an immunoprecipitated preparation is used; other proteins may also be pulled down in the same preparation and may actually be the active factor in this assay. (Extended fig 3 C/D). Results from Extended data 4 using anti-S6K antibody have not been included due to lack of specificity of this antibody. The authors have stated that Flag-Sesn2 was of mouse origin and Flag-WDR24 was of human origin, and that the cell lines used in Figures 2C and 2E, Extended Data Figures 2/3 were all human HEK293T. lim@mskcc.org Nature 2017 Only protein-protein interactions imex curation imex curation peng-2017-2 human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Peng M., Yin N., Li MO. Results from Amino acid binding assay have not been included as an immunoprecipitated preparation is used; other proteins may also be pulled down in the same preparation and may actually be the active factor in this assay. (Extended fig 3 C/D). Results from Extended data 4 using anti-S6K antibody have not been included due to lack of specificity of this antibody. The authors have stated that Flag-Sesn2 was of mouse origin and Flag-WDR24 was of human origin, and that the cell lines used in Figures 2C and 2E, Extended Data Figures 2/3 were all human HEK293T. lim@mskcc.org Nature 2017 Only protein-protein interactions imex curation imex curation peng-2017-3 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. molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Peng M., Yin N., Li MO. Results from Amino acid binding assay have not been included as an immunoprecipitated preparation is used; other proteins may also be pulled down in the same preparation and may actually be the active factor in this assay. (Extended fig 3 C/D). Results from Extended data 4 using anti-S6K antibody have not been included due to lack of specificity of this antibody. The authors have stated that Flag-Sesn2 was of mouse origin and Flag-WDR24 was of human origin, and that the cell lines used in Figures 2C and 2E, Extended Data Figures 2/3 were all human HEK293T. lim@mskcc.org Nature 2017 Only protein-protein interactions imex curation imex curation peng-2017-6 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. density sedimentation cosedimentation through density gradient density sedimentation western blot western blot Immuno blot Immuno blot Peng M., Yin N., Li MO. Results from Amino acid binding assay have not been included as an immunoprecipitated preparation is used; other proteins may also be pulled down in the same preparation and may actually be the active factor in this assay. (Extended fig 3 C/D). Results from Extended data 4 using anti-S6K antibody have not been included due to lack of specificity of this antibody. The authors have stated that Flag-Sesn2 was of mouse origin and Flag-WDR24 was of human origin, and that the cell lines used in Figures 2C and 2E, Extended Data Figures 2/3 were all human HEK293T. lim@mskcc.org Nature 2017 Only protein-protein interactions imex curation imex curation peng-2017-4 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. crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Peng M., Yin N., Li MO. Results from Amino acid binding assay have not been included as an immunoprecipitated preparation is used; other proteins may also be pulled down in the same preparation and may actually be the active factor in this assay. (Extended fig 3 C/D). Results from Extended data 4 using anti-S6K antibody have not been included due to lack of specificity of this antibody. Cells were cross linked with DSP (dithiobis(succinimidyl propionate) before IP with anti-WDR24. The authors have stated that Flag-Sesn2 was of mouse origin and Flag-WDR24 was of human origin, and that the cell lines used in Figures 2C and 2E, Extended Data Figures 2/3 were all human HEK293T. lim@mskcc.org Nature 2017 Only protein-protein interactions imex curation imex curation peng-2017-5 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 ms participant Identification by mass spectrometry ms participant Peng M., Yin N., Li MO. Results from Amino acid binding assay have not been included as an immunoprecipitated preparation is used; other proteins may also be pulled down in the same preparation and may actually be the active factor in this assay. (Extended fig 3 C/D). Results from Extended data 4 using anti-S6K antibody have not been included due to lack of specificity of this antibody. The authors have stated that Flag-Sesn2 was of mouse origin and Flag-WDR24 was of human origin, and that the cell lines used in Figures 2C and 2E, Extended Data Figures 2/3 were all human HEK293T. lim@mskcc.org Nature 2017 Only protein-protein interactions imex curation imex curation peng-2017-7 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. confocal microscopy confocal microscopy immunocytochemistry immunocytochemistry Peng M., Yin N., Li MO. Results from Amino acid binding assay have not been included as an immunoprecipitated preparation is used; other proteins may also be pulled down in the same preparation and may actually be the active factor in this assay. (Extended fig 3 C/D). Results from Extended data 4 using anti-S6K antibody have not been included due to lack of specificity of this antibody. The authors have stated that Flag-Sesn2 was of mouse origin and Flag-WDR24 was of human origin, and that the cell lines used in Figures 2C and 2E, Extended Data Figures 2/3 were all human HEK293T. lim@mskcc.org Nature 2017 Only protein-protein interactions imex curation imex curation wdr24_human GATOR complex protein WDR24 WDR24 C16orf21 WD repeat-containing protein 24 JFP7 protein protein human Homo sapiens Human MEKMSRVTTALGGSVLTGRTMHCHLDAPANAISVCRDAAQVVVAGRSIFKIYAIEEEQFVEKLNLRVGRKPSLNLSCADVVWHQMDENLLATAATNGVVVTWNLGRPSRNKQDQLFTEHKRTVNKVCFHPTEAHVLLSGSQDGFMKCFDLRRKDSVSTFSGQSESVRDVQFSIRDYFTFASTFENGNVQLWDIRRPDRCERMFTAHNGPVFCCDWHPEDRGWLATGGRDKMVKVWDMTTHRAKEMHCVQTIASVARVKWRPECRHHLATCSMMVDHNIYVWDVRRPFVPAAMFEEHRDVTTGIAWRHPHDPSFLLSGSKDSSLCQHLFRDASQPVERANPEGLCYGLFGDLAFAAKESLVAAESGRKPYTGDRRHPIFFKRKLDPAEPFAGLASSALSVFETEPGGGGMRWFVDTAERYALAGRPLAELCDHNAKVARELGRNQVAQTWTMLRIIYCSPGLVPTANLNHSVGKGGSCGLPLMNSFNLKDMAPGLGSETRLDRSKGDARSDTVLLDSSATLITNEDNEETEGSDVPADYLLGDVEGEEDELYLLDPEHAHPEDPECVLPQEAFPLRHEIVDTPPGPEHLQDKADSPHVSGSEADVASLAPVDSSFSLLSVSHALYDSRLPPDFFGVLVRDMLHFYAEQGDVQMAVSVLIVLGERVRKDIDEQTQEHWYTSYIDLLQRFRLWNVSNEVVKLSTSRAVSCLNQASTTLHVNCSHCKRPMSSRGWVCDRCHRCASMCAVCHHVVKGLFVWCQGCSHGGHLQHIMKWLEGSSHCPAGCGHLCEYS mio_human GATOR complex protein MIOS MIOS Missing oocyte meiosis regulator homolog protein protein human Homo sapiens Human MSGTKPDILWAPHHVDRFVVCDSELSLYHVESTVNSELKAGSLRLSEDSAATLLSINSDTPYMKCVAWYLNYDPECLLAVGQANGRVVLTSLGQDHNSKFKDLIGKEFVPKHARQCNTLAWNPLDSNWLAAGLDKHRADFSVLIWDICSKYTPDIVPMEKVKLSAGETETTLLVTKPLYELGQNDACLSLCWLPRDQKLLLAGMHRNLAIFDLRNTSQKMFVNTKAVQGVTVDPYFHDRVASFYEGQVAIWDLRKFEKPVLTLTEQPKPLTKVAWCPTRTGLLATLTRDSNIIRLYDMQHTPTPIGDETEPTIIERSVQPCDNYIASFAWHPTSQNRMIVVTPNRTMSDFTVFERISLAWSPITSLMWACGRHLYECTEEENDNSLEKDIATKMRLRALSRYGLDTEQVWRNHILAGNEDPQLKSLWYTLHFMKQYTEDMDQKSPGNKGSLVYAGIKSIVKSSLGMVESSRHNWSGLDKQSDIQNLNEERILALQLCGWIKKGTDVDVGPFLNSLVQEGEWERAAAVALFNLDIRRAIQILNEGASSEKGDLNLNVVAMALSGYTDEKNSLWREMCSTLRLQLNNPYLCVMFAFLTSETGSYDGVLYENKVAVRDRVAFACKFLSDTQLNRYIEKLTNEMKEAGNLEGILLTGLTKDGVDLMESYVDRTGDVQTASYCMLQGSPLDVLKDERVQYWIENYRNLLDAWRFWHKRAEFDIHRSKLDPSSKPLAQVFVSCNFCGKSISYSCSAVPHQGRGFSQYGVSGSPTKSKVTSCPGCRKPLPRCALCLINMGTPVSSCPGGTKSDEKVDLSKDKKLAQFNNWFTWCHNCRHGGHAGHMLSWFRDHAECPVSACTCKCMQLDTTGNLVPAETVQP nprl2_human GATOR complex protein NPRL2 NPRL2 Gene 21 protein TUSC4 Tumor suppressor candidate 4 Nitrogen permease regulator 2-like protein protein protein human Homo sapiens Human MGSGCRIECIFFSEFHPTLGPKITYQVPEDFISRELFDTVQVYIITKPELQNKLITVTAMEKKLIGCPVCIEHKKYSRNALLFNLGFVCDAQAKTCALEPIVKKLAGYLTTLELESSFVSMEESKQKLVPIMTILLEELNASGRCTLPIDESNTIHLKVIEQRPDPPVAQEYDVPVFTKDKEDFFNSQWDLTTQQILPYIDGFRHIQKISAEADVELNLVRIAIQNLLYYGVVTLVSILQYSNVYCPTPKVQDLVDDKSLQEACLSYVTKQGHKRASLRDVFQLYCSLSPGTTVRDLIGRHPQQLQHVDERKLIQFGLMKNLIRRLQKYPVRVTREEQSHPARLYTGCHSYDEICCKTGMSYHELDERLENDPNIIICWK nprl3_human GATOR complex protein NPRL3 MARE NPRL3 C16orf35 CGTHBA Alpha-globin regulatory element-containing gene protein Protein CGTHBA -14 gene protein Nitrogen permease regulator 3-like protein protein protein human Homo sapiens Human MRDNTSPISVILVSSGSRGNKLLFRYPFQRSQEHPASQTSKPRSRYAASNTGDHADEQDGDSRFSDVILATILATKSEMCGQKFELKIDNVRFVGHPTLLQHALGQISKTDPSPKREAPTMILFNVVFALRANADPSVINCLHNLSRRIATVLQHEERRCQYLTREAKLILALQDEVSAMADGNEGPQSPFHHILPKCKLARDLKEAYDSLCTSGVVRLHINSWLEVSFCLPHKIHYAASSLIPPEAIERSLKAIRPYHALLLLSDEKSLLGELPIDCSPALVRVIKTTSAVKNLQQLAQDADLALLQVFQLAAHLVYWGKAIIIYPLCENNVYMLSPNASVCLYSPLAEQFSHQFPSHDLPSVLAKFSLPVSLSEFRNPLAPAVQETQLIQMVVWMLQRRLLIQLHTYVCLMASPSEEEPRPREDDVPFTARVGGRSLSTPNALSFGSPTSSDDMTLTSPSMDNSSAELLPSGDSPLNQRMTENLLASLSEHERAAILSVPAAQNPEDLRMFARLLHYFRGRHHLEEIMYNENTRRSQLLMLFDKFRSVLVVTTHEDPVIAVFQALLP szt2_human KICSTOR complex protein SZT2 SZT2 C1orf84 KIAA0467 Seizure threshold 2 protein homolog protein protein human Homo sapiens Human MASERPEPEVEEAGQVFLLMKKDYRISRNVRLAWFLSHLHQTVQATPQEMLLQSEQELEVLSVLPPGWQPDEPVVPRPFLLVPSTRVTFLAWQYRFVIELDLSPSTGIVDDSTGEILFDEVFHALSRCLGGLLRPFRVPGSCIDFQPEIYVTIQAYSSIIGLQSHQVLVQGCLLDPSQREVFLQQIYEQLCLFEDKVATMLQQQYDPQSQAEDQSPDSGDLLGRKVGVSMVTADLGLVSMIRQGILALQLLPSNSSAGIIVITDGVTSVPDVAVCETLLNQLRSGTVACSFVQVGGVYSYDCSFGHVPNVELMKFIAMATFGSYLSTCPEPEPGNLGLTVYHRAFLLYSFLRSGEALNPEYYCGSQHRLFNEHLVSASSNPALALRRKKHTEKEVPADLVSTVSVRLREGYSVREVTLAKGGSQLEVKLVLLWKHNMRIEYVAMAPWPLEPEGPRVTRVEVTMEGGYDILHDVSCALRQPIRSLYRTHVIRRFWNTLQSINQTDQMLAHLQSFSSVPEHFTLPDSTKSGVPLFYIPPGSTTPVLSLQPSGSDSSHAQFAAYWKPVLSMDANSWQRWLHMHRLVLILEHDTPIPKHLHTPGSNGRYSTIQCRISHSSLTSLLRDWSSFVLVEGYSYVKLLSSAPDQPPNSFYMVRIISKAPCMVLRLGFPIGTPAPARHKIVSGLREEILRLRFPHRVQSKEPTPKVKRKGLGGAGGGSSPSKSPPVLGPQQALSDRPCLVVLHKPLDKLLIRYEKLPLDYRAPFLLTLEPPGPLPLVSGRSASSSLASLSRYLYHQRWLWSVPSGLAPALPLSAIAQLLSILTEVRLSEGFHFACSGEGIINMVLELPIQNEPPGQAAAEEKHTCVVQYILFPPHSTSTKDSFSTDDDNDVEVEALEGDSELNLVTEVWVEPQYGRVGPGPGIWKHLQDLTYSEIPQALHPRDAACIGSMLSFEYLIQLCQSKEWGPLPPEPRVSDGLDQGGDTCVHEIPFHFDLMGLLPQCQQLQMFFLLLAREPEGVPFAEGSCPANDMVLCLLHSCLGQELSDREIPLTPVDQAAFLSEVLRRTCHVPGAEGPLLGVHGIPKEQAVGSTQATGDSAFTSLSVGLPETLKPLISAQPPQWRCYARLVNPQHVFLTFLPATFSDVQRLAACGLEGPPQEETKPKFGDWSGAPSLKDLGGTGIKATKSHVPVLSVTLASDNAQNQGELSPPFRRDLQAYAGRQASQTESADGPRTRCPVYIYSCSLEALREQMVGMQPPQAPRDLIFRTQFLDHPSPSSAWMEPRYKEAANHCALLQEHAQRCYVRGLFRSLQQAQSVTSQDLLTAVDACEELLQEIDITPFLLALCGHTWGLPHAPPSPGPLSPGPFSSSMEEGAEPRERAILASESSIETEDLSEPEFQSTRVPGIPDPGPEISLTDVCQLRGEAHGALHSVIQEKFLEISRLHFRTVPSNPHYFFYCPPSSRREDEGPRDTVDRKISDLEFSEAELMGEEGDTSACCVVTESDPELEVEYRESRESDLGPAGLDSASLSDVDTVNPDEDSFSILGGDSPTGPESFLHDLPPLFLHLTCSVRLRGQHSSVPVCSLPTCLGQVLSSLEGPPVGGRVPLRDLSVTLDVFMLTLPLEVELPTASDPQHHRSTSESSASFPRSPGQPSSLRSDDGLGPPLPPPEEERHPGLSNLATPHRLAIETTMNEIRWLLEDEMVGALRRGGIPQSPALHRAAAHIHSSPGRSTCLRQTLPLSFVFGPERSLTQFKEEFRRLHLPGHVLLEDPDSGFFFVAAGQQPGGSHGEPSSAAWAWHSHEDRAEGIEGETLTASPQAPGSPEDSEGVPLISLPRVPQGGSQPGPSRGLSLMSSQGSVDSDHLGYDGGSSGSDSEGPNDTLGEKAPFTLRTPPGPAPPQPSLSGLPGPCLPDFWLIVRVLQDRVEVYAHARSLIREDGGPGTECRHLQQLLVRRVGEICREVNQRLLLQDLHDSHVCNSLLVAESEEDLWRSETPFHSRQRAPLPSDDYAADESCAPRGYLAATMQFVPGHFSCDVVWGTVIRVHSRLKMGPSMGVSRAIQALRSVLNAFSVVNRKNMFVYQERATKAVYYLRLLETSCSDRPWKGDALPPSLALSRSQEPIYSEEASGPRSPLDMVSSRSSDAARPVGQVDRHIQLLVHGVGQAGPEITDELVRVLCRRLDEATLDVITVMLVRNCKLTPADVEFIQPPGSLPSEVLHLALPTSCRPWLPALAWYLRQNLLIFLHSPKYTDSNSRNHFQHPLPPQGGLPDLDIYLYNKPGGQGTGGKGVACITLAFVDEGGAPLSLALWPPSSPGPPDPLREEEFEQLTQVIRCPVVVDSSSAQNGAPRLRLDVWEKGNISIVQLEEKLRGAARQALADAIIELQLLPASLCTEDTPTGSLRNGSLETKSSAGRASTFPPAPVPGEPVTPPSKAGRRSFWDMLSKTECGDLGSPKTTDDIVLDRPEDTRGRRRHKTESVRTPGGAERAPGSDSGAQRQKRRTTQLEEGEVGTLHPVFARVAQRWMEFMVQIGCASVSRSSAHMVSRFLLPSILSEFTALVTSMAGDTSVRIFEQHLGSEPEIFGPCSPGQLGPSPRPAAERHLLLLGRNFLQWRRPTQQAAKAMQRFEPGGDGSSGRNAPRQRLLLLEVVDKKLQLLTYNWAPDLGAALGRALVRLVQWQNARAHLIFCLLSQKLGLFHHYGQLDFPVRDEKEPNPFLLPTMEVETLIRSASPPLSREQGRLSGSSRGGGPLPLDTFPFDEALRDITAARPSSVLGPVPRPPDPVTYHGQQFLEIKMAERRELERQMKMENLFVTWQQRSTPATMPISAGELETLKQSSRLVHYCATAMLFDPAAWLHGPPETSGPPDGQRRHRPESGSGSREAPTSCESLDVSPPGAREEPWLKELSLAFLQQYVQYLQSIGFVLVPLRPPSPARSTSRPRAMAILGTEGRGSFSCPKTKTDGSPKSTSSPVTTYHLQRALPGGIILMELAFQGCYFCVKQFALECSRIPMGQAVNSQLSMLFTEECDKVRDLMHVHSFSYDFHLRLVHQHVLGAHLVLRHGYHLTTFLRHFLAHHPDGPHFGRNHIYQGTLELPTPLIAAHQLYNYVADHASSYHMKPLRMARPGGPEHNEYALVSAWHSSGSYLDSEGLRHQDDFDVSLLVCHCAAPFEEQGEAERHVLRLQFFVVLTSQRELFPRLTADMRRFRKPPRLPPEPEAPGSSAGSPGEASGLILAPGPAPLFPPLAAEVGMARARLAQLVRLAGGHCRRDTLWKRLFLLEPPGPDRLRLGGRLALAELEELLEAVHAKSIGDIDPQLDCFLSMTVSWYQSLIKVLLSRFPQSCRHFQSPDLGTQYLVVLNQKFTDCFVLVFLDSHLGKTSLTVVFREPFPVQPQDSESPPAQLVSTYHHLESVINTACFTLWTRLL wdr59_human GATOR complex protein WDR59 WDR59 KIAA1923 WD repeat-containing protein 59 FP977 protein protein human Homo sapiens Human MAARWSSENVVVEFRDSQATAMSVDCLGQHAVLSGRRFLYIVNLDAPFEGHRKISRQSKWDIGAVQWNPHDSFAHYFAASSNQRVDLYKWKDGSGEVGTTLQGHTRVISDLDWAVFEPDLLVTSSVDTYIYIWDIKDTRKPTVALSAVAGASQVKWNKKNANCLATSHDGDVRIWDKRKPSTAVEYLAAHLSKIHGLDWHPDSEHILATSSQDNSVKFWDYRQPRKYLNILPCQVPVWKARYTPFSNGLVTVMVPQLRRENSLLLWNVFDLNTPVHTFVGHDDVVLEFQWRKQKEGSKDYQLVTWSRDQTLRMWRVDSQMQRLCANDILDGVDEFIESISLLPEPEKTLHTEDTDHQHTASHGEEEALKEDPPRNLLEERKSDQLGLPQTLQQEFSLINVQIRNVNVEMDAADRSCTVSVHCSNHRVKMLVKFPAQYPNNAAPSFQFINPTTITSTMKAKLLKILKDTALQKVKRGQSCLEPCLRQLVSCLESFVNQEDSASSNPFALPNSVTPPLPTFARVTTAYGSYQDANIPFPRTSGARFCGAGYLVYFTRPMTMHRAVSPTEPTPRSLSALSAYHTGLIAPMKIRTEAPGNLRLYSGSPTRSEKEQVSISSFYYKERKSRRWKSKREGSDSGNRQIKAAGKVIIQDIACLLPVHKSLGELYILNVNDIQETCQKNAASALLVGRKDLVQVWSLATVATDLCLGPKSDPDLETPWARHPFGRQLLESLLAHYCRLRDVQTLAMLCSVFEAQSRPQGLPNPFGPFPNRSSNLVVSHSRYPSFTSSGSCSSMSDPGLNTGGWNIAGREAEHLSSPWGESSPEELRFGSLTYSDPRERERDQHDKNKRLLDPANTQQFDDFKKCYGEILYRWGLREKRAEVLKFVSCPPDPHKGIEFGVYCSHCRSEVRGTQCAICKGFTFQCAICHVAVRGSSNFCLTCGHGGHTSHMMEWFRTQEVCPTGCGCHCLLESTF sec13_human Protein SEC13 homolog SEC13 D3S1231E SEC13L1 SEC13R SEC13-related protein SEC13-like protein 1 GATOR complex protein SEC13 SEC13A protein protein human Homo sapiens Human MVSVINTVDTSHEDMIHDAQMDYYGTRLATCSSDRSVKIFDVRNGGQILIADLRGHEGPVWQVAWAHPMYGNILASCSYDRKVIIWREENGTWEKSHEHAGHDSSVNSVCWAPHDYGLILACGSSDGAISLLTYTGEGQWEVKKINNAHTIGCNAVSWAPAVVPGSLIDHPSGQKPNYIKRFASGGCDNLIKLWKEEEDGQWKEEQKLEAHSDWVRDVAWAPSIGLPTSTIASCSQDGRVFIWTCDDASSNTWSPKLLHKFNDVVWHVSWSITANILAVSGGDNKVTLWKESVDGQWVCISDVNKGQGSVSASVTEGQQNEQ seh1_human Nucleoporin SEH1 Nup107-160 subcomplex subunit SEH1 SEH1L SEC13L SEH1 SEC13-like protein GATOR complex protein SEH1 protein protein human Homo sapiens Human MFVARSIAADHKDLIHDVSFDFHGRRMATCSSDQSVKVWDKSESGDWHCTASWKTHSGSVWRVTWAHPEFGQVLASCSFDRTAAVWEEIVGESNDKLRGQSHWVKRTTLVDSRTSVTDVKFAPKHMGLMLATCSADGIVRIYEAPDVMNLSQWSLQHEISCKLSCSCISWNPSSSRAHSPMIAVGSDDSSPNAMAKVQIFEYNENTRKYAKAETLMTVTDPVHDIAFAPNLGRSFHILAIATKDVRIFTLKPVRKELTSSGGPTKFEIHIVAQFDNHNSQVWRVSWNITGTVLASSGDDGCVRLWKANYMDNWKCTGILKGNGSPVNGSSQQGTSNPSLGSTIPSLQNSLNGSSAGRKHS rragb_human Ras-related GTP-binding protein B RRAGB protein protein human Homo sapiens Human MEESDSEKTTEKENLGPRMDPPLGEPEGSLGWVLPNTAMKKKVLLMGKSGSGKTSMRSIIFANYIARDTRRLGATILDRIHSLQINSSLSTYSLVDSVGNTKTFDVEHSHVRFLGNLVLNLWDCGGQDTFMENYFTSQRDNIFRNVEVLIYVFDVESRELEKDMHYYQSCLEAILQNSPDAKIFCLVHKMDLVQEDQRDLIFKEREEDLRRLSRPLECSCFRTSIWDETLYKAWSSIVYQLIPNVQQLEMNLRNFAEIIEADEVLLFERATFLVISHYQCKEQRDAHRFEKISNIIKQFKLSCSKLAASFQSMEVRNSNFAAFIDIFTSNTYVMVVMSDPSIPSAATLINIRNARKHFEKLERVDGPKQCLLMR rragc_human Ras-related GTP-binding protein C GTPase-interacting protein 2 TIB929 RRAGC protein protein human Homo sapiens Human MSLQYGAEETPLAGSYGAADSFPKDFGYGVEEEEEEAAAAGGGVGAGAGGGCGPGGADSSKPRILLMGLRRSGKSSIQKVVFHKMSPNETLFLESTNKIYKDDISNSSFVNFQIWDFPGQMDFFDPTFDYEMIFRGTGALIYVIDAQDDYMEALTRLHITVSKAYKVNPDMNFEVFIHKVDGLSDDHKIETQRDIHQRANDDLADAGLEKLHLSFYLTSIYDHSIFEAFSKVVQKLIPQLPTLENLLNIFISNSGIEKAFLFDVVSKIYIATDSSPVDMQSYELCCDMIDVVIDVSCIYGLKEDGSGSAYDKESMAIIKLNNTTVLYLKEVTKFLALVCILREESFERKGLIDYNFHCFRKAIHEVFEVGVTSHRSCGHQTSASSLKALTHNGTPRNAI o75140-4 GATOR complex protein DEPDC5 DEPDC5 KIAA0645 DEP domain-containing protein 5 protein protein human Homo sapiens Human MRTTKVYKLVIHKKGFGGSDDELVVNPKVFPHIKLGDIVEIAHPNDEYSPLLLQVKSLKEDLQKETISVDQTVTQVFRLRPYQDVYVNVVDPKDVTLDLVELTFKDQYIGRGDMWRLKKSLVSTCAYITQKVEFAGIRAQAGELWVKNEKVMCGYISEDTRVVFRSTSAMVYIFIQMSCEMWDFDIYGDLYFEKAVNGFLADLFTKWKEKNCSHEVTVVLFSRTFYDAKSVDEFPEINRASIRQDHKGRFYEDFYKVVVQNERREEWTSLLVTIKKLFIQYPVLVRLEQAEGFPQGDNSTSAQGNYLEAINLSFNVFDKHYINRNFDRTGQMSVVITPGVGVFEVDRLLMILTKQRMIDNGIGVDLVCMGEQPLHAVPLFKLHNRSAPRDSRLGDDYNIPHWINHSFYTSKSQLFCNSFTPRIKLAGKKPASEKAKNGRDTSLGSPKESENALPIQVDYDAYDAQVFRLPGPSRAQCLTTCRSVRERESHSRKSASSCDVSSSPSLPSRTLPTEEVRSQASDDSSLGKSANILMIPHPHLHQYEVSSSLGYTSTRDVLENMMEPPQRDSSAPGRFHVGSAESMLHVRPGGYTPQRALINPFAPSRMPMKLTSNRRRWMHTFPVGPSGEAIQIHHQTRQNMAELQGSGQRDPTHSSAELLELAYHEAAGRHSNSRQPGDGMSFLNFSGTEELSVGLLSNSGAGMNPRTQNKDSLEDSVSTSPDPILTLSAPPVVPGFCCTVGVDWKSLTTPACLPLTTDYFPDRQGLQNDYTEGCYDLLPEADIDRRDEDGVQMTAQQVFEEFICQRLMQGYQIIVQPKTQKPNPAVPPPLSSSPLYSRGLVSRNRPEEEDQYWLSMGRTFHKVTLKDKMITVTRYLPKYPYESAQIHYTYSLCPSHSDSEFVSCWVEFSHERLEEYKWNYLDQYICSAGSEDFSLIESLKFWRTRFLLLPACVTATKRITEGEAHCDIYGDRPRADEDEWQLLDGFVRFVEGLNRIRRRHRSDRMMRKGTAMKGLQMTGPISTHSLESTAPPVGKKGTSALSALLEMEASQKCLGEQQAAVHGGKSSAQSAESSSVAMTPTYMDSPRKVSVDQTATPMLDGTSLGICTGQSMDRGNSQTFGNSQNIGEQGYSSTNSSDSSSQQLVASSLTSSSTLTEILEAMKHPSTGVQLLSEQKGLSPYCFISAEVVHWLVNHVEGIQTQAMAIDIMQKMLEEQLITHASGEAWRTFIYGFYFYKIVTDKEPDRVAMQQPATTWHTAGVDDFASFQRKWFEVAFVAEELVHSEIPAFLLPWLPSRPASYASRHSSFSRSFGGRSQAAALLAATVPEQRTVTLDVDVNNRTDRLEWCSCYYHGNFSLNAAFEIKLHWMAVTAAVLFEMVQGWHRKATSCGFLLVPVLEGPFALPSYLYGDPLRAQLFIPLNISCLLKEGSEHLFDSFEPETYWDRMHLFQEAIAHRFGFVQDKYSASAFNFPAENKPQYIHVTGTVFLQLPYSKRKFSGQQRRRRNSTSSTNQNMFCEERVGYNWAYNTMLTKTWRSSATGDEKFADRLLKDFTDFCINRDNRLVTFWTSCLEKMHASAP No experimental confirmation available. sesn2_human Sestrin-2 SESN2 SEST2 Hi95 Hypoxia-induced gene protein protein human Homo sapiens Human MIVADSECRAELKDYLRFAPGGVGDSGPGEEQRESRARRGPRGPSAFIPVEEVLREGAESLEQHLGLEALMSSGRVDNLAVVMGLHPDYFTSFWRLHYLLLHTDGPLASSWRHYIAIMAAARHQCSYLVGSHMAEFLQTGGDPEWLLGLHRAPEKLRKLSEINKLLAHRPWLITKEHIQALLKTGEHTWSLAELIQALVLLTHCHSLSSFVFGCGILPEGDADGSPAPQAPTPPSEQSSPPSRDPLNNSGGFESARDVEALMERMQQLQESLLRDEGTSQEEMESRFELEKSESLLVTPSADILEPSPHPDMLCFVEDPTFGYEDFTRRGAQAPPTFRAQDYTWEDHGYSLIQRLYPEGGQLLDEKFQAAYSLTYNTIAMHSGVDTSVLRRAIWNYIHCVFGIRYDDYDYGEVNQLLERNLKVYIKTVACYPEKTTRRMYNLFWRHFRHSEKVHVNLLLLEARMQAALLYALRAITRYMT lamp1_human Lysosome-associated membrane glycoprotein 1 CD107 antigen-like family member A LAMP1 protein protein human Homo sapiens Human MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMANFSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNFTRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHMNNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKYNVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSEGTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNAEEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIVLIAYLVGRKRSHAGYQTI depd5_human GATOR complex protein DEPDC5 DEPDC5 DEP domain-containing protein 5 KIAA0645 protein protein human Homo sapiens Human MRTTKVYKLVIHKKGFGGSDDELVVNPKVFPHIKLGDIVEIAHPNDEYSPLLLQVKSLKEDLQKETISVDQTVTQVFRLRPYQDVYVNVVDPKDVTLDLVELTFKDQYIGRGDMWRLKKSLVSTCAYITQKVEFAGIRAQAGELWVKNEKVMCGYISEDTRVVFRSTSAMVYIFIQMSCEMWDFDIYGDLYFEKAVNGFLADLFTKWKEKNCSHEVTVVLFSRTFYDAKSVDEFPEINRASIRQDHKGRFYEDFYKVVVQNERREEWTSLLVTIKKLFIQYPVLVRLEQAEGFPQGDNSTSAQGNYLEAINLSFNVFDKHYINRNFDRTGQMSVVITPGVGVFEVDRLLMILTKQRMIDNGIGVDLVCMGEQPLHAVPLFKLHNRSAPRDSRLGDDYNIPHWINHSFYTSKSQLFCNSFTPRIKLAGKKPASEKAKNGRDTSLGSPKESENALPIQVDYDAYDAQVFRLPGPSRAQCLTTCRSVRERESHSRKSASSCDVSSSPSLPSRTLPTEEVRSQASDDSSLGKSANILMIPHPHLHQYEVSSSLGYTSTRDVLENMMEPPQRDSSAPGRFHVGSAESMLHVRPGGYTPQRALINPFAPSRMPMKLTSNRRRWMHTFPVGPSGEAIQIHHQTRQNMAELQGSGQRDPTHSSAELLELAYHEAAGRHSNSRQPGDGMSFLNFSGTEELSVGLLSNSGAGMNPRTQNKDSLEDSVSTSPDPILTLSAPPVVPGFCCTVGVDWKSLTTPACLPLTTDYFPDRQGLQNDYTEGCYDLLPEADIDRRDEDGVQMTAQQVFEEFICQRLMQGYQIIVQPKTQKPNPAVPPPLSSSPLYSRGLVSRNRPEEEDQYWLSMGRTFHKVTLKDKMITVTRYLPKYPYESAQIHYTYSLCPSHSDSEFVSCWVEFSHERLEEYKWNYLDQYICSAGSEDFSLIESLKFWRTRFLLLPACVTATKRITEGEAHCDIYGDRPRADEDEWQLLDGFVRFVEGLNRIRRRHRSDRMMRKGTAMKGLQMTGPISTHSLESTAPPVGKKGTSALSALLEMEASQKCLGEQQAAVHGGKSSAQSAESSSVAMTPTYMDSPRKDGAFFMEFVRSPRTASSAFYPQVSVDQTATPMLDGTSLGICTGQSMDRGNSQTFGNSQNIGEQGYSSTNSSDSSSQQLVASSLTSSSTLTEILEAMKHPSTGVQLLSEQKGLSPYCFISAEVVHWLVNHVEGIQTQAMAIDIMQKMLEEQLITHASGEAWRTFIYGFYFYKIVTDKEPDRVAMQQPATTWHTAGVDDFASFQRKWFEVAFVAEELVHSEIPAFLLPWLPSRPASYASRHSSFSRSFGGRSQAAALLAATVPEQRTVTLDVDVNNRTDRLEWCSCYYHGNFSLNAAFEIKLHWMAVTAAVLFEMVQGWHRKATSCGFLLVPVLEGPFALPSYLYGDPLRAQLFIPLNISCLLKEGSEHLFDSFEPETYWDRMHLFQEAIAHRFGFVQDKYSASAFNFPAENKPQYIHVTGTVFLQLPYSKRKFSGQQRRRRNSTSSTNQNMFCEERVGYNWAYNTMLTKTWRSSATGDEKFADRLLKDFTDFCINRDNRLVTFWTSCLEKMHASAP lamp2_human Lysosome-associated membrane glycoprotein 2 LAMP2 CD107 antigen-like family member B LGP-96 protein protein human Homo sapiens Human MVCFRLFPVPGSGLVLVCLVLGAVRSYALELNLTDSENATCLYAKWQMNFTVRYETTNKTYKTVTISDHGTVTYNGSICGDDQNGPKIAVQFGPGFSWIANFTKAASTYSIDSVSFSYNTGDNTTFPDAEDKGILTVDELLAIRIPLNDLFRCNSLSTLEKNDVVQHYWDVLVQAFVQNGTVSTNEFLCDKDKTSTVAPTIHTTVPSPTTTPTPKEKPEAGTYSVNNGNDTCLLATMGLQLNITQDKVASVININPNTTHSTGSCRSHTALLRLNSSTIKYLDFVFAVKNENRFYLKEVNISMYLVNGSVFSIANNNLSYWDAPLGSSYMCNKEQTVSVSGAFQINTFDLRVQPFNVTQGKYSTAQDCSADDDNFLVPIAVGAALAGVLILVLLAYFIGLKHHHAGYEQF szt2-mios-1 3338709 N/A 3338716 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338717 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338718 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338719 ancillary ancillary prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338720 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed infection infection Q5T011:p.Ile1938Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 3338721 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338722 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338723 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Figure 2A/B/F, Extended Data figure 3 A/B,4 In the absence of NPRL3 this entire interaction was not seen, and was substantially diminished in the absence of WDR59, WDR24 and MIOS. The SZT2–GATOR interaction was not modulated by amino acids, and the stable SOG complex did not dissociate during washes with 1.5 M sodium chloride. depdc5-mios-1 3338709 N/A 3338724 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338725 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338717 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338718 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338719 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338726 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed infection infection n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false association association false false false Figure 2A-2 positive control szt2-mios-2 3338709 N/A 3338718 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338719 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338720 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed infection infection Q5T011:p.Ile1938Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false association association false false false Figure 2F Weak interaction in the absence of WDR59, WDR24 and MIOS. szt2-nprl2-1 3338709 N/A 3338719 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338720 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed infection infection Q5T011:p.Ile1938Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Extended Data 4 szt2-nprl3-1 3338709 N/A 3338716 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338720 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed infection infection region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q5T011:p.Ile1938Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Extended Data 4-D Deletion of a small portion (amino acids 1,083–1,189) within the GATOR1-binding region (amino acids 935–1,734) created a SZT2 mutant, that had minimal binding to NPRL3. 123456qwerty-1 3338709 N/A 3338721 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338727 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338717 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role bait bait engineered engineered 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 Extended Data 3E Interaction seen when cells stably over-expressing Flag–WDR24 were deprived of amino acids. The interaction of SESN2 with GATOR2 (WDR24/59/MIOS) was unchanged in the absence of SZT2. wdr24-mios-9 3338709 N/A 3338721 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338717 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role bait bait engineered engineered 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 Extended Data 3E-2 Interaction seen when cells stably over-expressing Flag–WDR24 were re-stimulated with amino acids after prior deprivation. wdr24-mios-10 3338709 N/A 3338719 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338720 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed infection infection region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3338721 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338717 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Extended Data 3F SESN2 overexpression did not affect the interaction between SZT2, GATOR1 and GATOR2. depdc5-mios-2 3338710 N/A 3338720 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed infection infection Q5T011:p.Ile1938Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 3338727 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338721 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338717 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338718 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338719 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Figure 2B-3 SESN2 pulled down GATOR2, GATOR1 and SZT2. depdc5-mios-3 3338710 N/A 3338716 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338717 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338721 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Figure 2B/D Endogenous WDR24 co-immunoprecipitated efficiently with WDR59 and MIOS, but not with NPRL2 or NPRL3. wdr24-mios-7 3338710 N/A 3338718 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338719 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Figure 2B-2 NPRL3 pulled down NPRL2, but not WDR59, WDR24 or MIOS. wdr24-mios-8 3338710 N/A 3338720 unspecified role unspecified role ancillary ancillary physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338719 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338721 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Figure 2D WDR24 pulled down NPRL3 only in SZT2-containing SEC fractions (13-20). sesn2-mios-3 3338711 N/A 3338721 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Figure 2C Interaction in absence of Flag-SZT2 transfection, higher molecular weight bands, fractions ~13-20. sesn2-mios-4 3338711 N/A 3338720 anti tag western anti tag western blot anti tag western unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed cell lysate cell lysate infection infection Q5T011:p.Ile1938Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 3338721 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338718 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338719 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Figure 2C Interaction seen in Flag-SZT2 transfected cells, higher molecular weight fractions only (13-20). nprl2-nprl3-1 3338711 N/A 3338721 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338718 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338719 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Figure 2Ci Interaction seen in non-Flag-SZT2 transfected cells, lower molecular weight bands, fractions ~23-28. nprl2-nprl3-4 3338711 N/A 3338720 anti tag western anti tag western blot anti tag western unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed partially purified partially purified infection infection Q5T011:p.Ile1938Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 3338721 unspecified role unspecified role neutral component neutral component partially purified partially purified N/A 3338716 unspecified role unspecified role neutral component neutral component partially purified partially purified N/A 3338718 unspecified role unspecified role neutral component neutral component partially purified partially purified N/A 3338719 unspecified role unspecified role neutral component neutral component partially purified partially purified N/A 3338723 unspecified role unspecified role neutral component neutral component partially purified partially purified N/A 3338722 unspecified role unspecified role neutral component neutral component partially purified partially purified N/A 3338717 unspecified role unspecified role neutral component neutral component partially purified partially purified association association false false false Extended Data Figure 2 F/G Fraction containing (purified) SOG complex nprl2-nprl3-5 3338712 N/A 3338720 anti tag western anti tag western blot anti tag western unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338721 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338718 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338719 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level colocalization colocalization false false false Extended Data Figure 2H Interaction seen in Sucrose fractions1-5 inclusive, in both the presence and absence of SZT2. nprl2-nprl3-6 3338712 N/A 3338728 unspecified role unspecified role neutral component neutral component cell lysate cell lysate N/A 3338720 anti tag western anti tag western blot anti tag western unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338721 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338719 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338718 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level colocalization colocalization false false false Extended Data Figure 2Hi Interaction seen in Sucrose fractions 7-17 inclusive, in the presence of SZT2. lamp1-nprl2-1 3338712 N/A 3338728 unspecified role unspecified role neutral component neutral component cell lysate cell lysate N/A 3338720 anti tag western anti tag western blot anti tag western unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338721 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level colocalization colocalization false false false Extended Data figure 2Hii Interaction seen in Sucrose fractions 9-17 inclusive, in the absence of SZT2, and also fractions 19-21 in presence and absence of SZT2, except for fraction 21 where Lamp1 is not detected in the presence of SZT2. nprl2-nprl3-2 3338713 N/A 3338720 ancillary ancillary prey prey N/A 3338721 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338718 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338719 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Figure 2E The presence of SZT2 was required to bind NPRL2/3. nprl2-nprl3-3 3338713 N/A 3338721 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338716 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Figure 2Ei Interaction in the absence of SZT2. szt2-mios-3 3338714 N/A 3338723 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338722 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338729 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338717 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338718 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338719 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3338720 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed infection infection Q5T011:p.Ile1938Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 3338721 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Extended Data Figure 2A lamp2-szt2-1 3338715 N/A 3338721 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell N/A 3338730 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell N/A 3338720 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed infection infection fixed cell fixed cell Q5T011:p.Ile1938Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false colocalization colocalization false false false Extended Data 5C