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/ The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The EMBO journal 2016 imex curation Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. k.de_vos@sheffield.ac.uk 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-sh_sy5y Homo sapiens SH-SY5Y cell line sh_sy5y Human neuroblastoma cells fluorescence imaging fluorescence microscopy fluorescence imaging immunocytochemistry immunocytochemistry Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. k.de_vos@sheffield.ac.uk Accepted 2018-JUN-19 AT 13:01 BST AT 13:01 BST by PPORRAS The EMBO journal 2016 Only protein-protein interactions imex curation imex curation added figure 6 - two source-statements; added- a longer comment for Figs 3D/E; The other expt deleted and source text added.; added source text - lots, and altered Fig legend; added; added The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The EMBO journal 2016 imex curation Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. k.de_vos@sheffield.ac.uk 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. for us, I see no difference between Fig 3 PLA and EV2 PLA? k.de_vos@sheffield.ac.uk Accepted 2018-JUN-19 AT 13:01 BST AT 13:01 BST by PPORRAS The EMBO journal 2016 Only protein-protein interactions imex curation imex curation added figure 6 - two source-statements; added- a longer comment for Figs 3D/E; The other expt deleted and source text added.; added source text - lots, and altered Fig legend; added; added The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The EMBO journal 2016 imex curation Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. k.de_vos@sheffield.ac.uk 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. k.de_vos@sheffield.ac.uk Accepted 2018-JUN-19 AT 13:01 BST AT 13:01 BST by PPORRAS The EMBO journal 2016 Only protein-protein interactions imex curation imex curation added figure 6 - two source-statements; added- a longer comment for Figs 3D/E; The other expt deleted and source text added.; added source text - lots, and altered Fig legend; added; added The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The EMBO journal 2016 imex curation Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. k.de_vos@sheffield.ac.uk 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. k.de_vos@sheffield.ac.uk Accepted 2018-JUN-19 AT 13:01 BST AT 13:01 BST by PPORRAS The EMBO journal 2016 Only protein-protein interactions imex curation imex curation added figure 6 - two source-statements; added- a longer comment for Figs 3D/E; The other expt deleted and source text added.; added source text - lots, and altered Fig legend; added; added The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The EMBO journal 2016 imex curation Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. k.de_vos@sheffield.ac.uk 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. Fig3A : Isoforms of C9orf72:I could have added one interaction rather than 2 - with "sufficient to bind" aas 1-221- but as aa 222 is different for both isoforms I added both.?? k.de_vos@sheffield.ac.uk Accepted 2018-JUN-19 AT 13:01 BST AT 13:01 BST by PPORRAS The EMBO journal 2016 Only protein-protein interactions imex curation imex curation added figure 6 - two source-statements; added- a longer comment for Figs 3D/E; The other expt deleted and source text added.; added source text - lots, and altered Fig legend; added; added The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The EMBO journal 2016 imex curation Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. k.de_vos@sheffield.ac.uk 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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. pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. Figure 7A: there is NO red dots for PLA visible in presence of C9orf72 siRNA - so have added "no-uniport-update" annotatation to other expt - E-9 (binary interaction) , and added C9orf72 as an "ancillary" here . For the "colocalisations" I can see some apparent co-loc visible even in the presence of relevant siRNA - so have just added the siRNA as "antagonist". k.de_vos@sheffield.ac.uk Accepted 2018-JUN-19 AT 13:01 BST AT 13:01 BST by PPORRAS The EMBO journal 2016 Only protein-protein interactions imex curation imex curation added figure 6 - two source-statements; added- a longer comment for Figs 3D/E; The other expt deleted and source text added.; added source text - lots, and altered Fig legend; added; added The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The EMBO journal 2016 imex curation Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. k.de_vos@sheffield.ac.uk 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 fluorescence imaging fluorescence microscopy fluorescence imaging immunocytochemistry immunocytochemistry Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. I think that Fig 4D/5 shows colocalisation, even if they are not trying to demonstrate this. Omited Fig 4C as i do not think co-loc is shown with mCerulean‐tagged constructs. Also Figure 7B - in text- specifically mentions "mCherry‐FIP200 and Rab1a(Q70L) appeared punctate and co‐localized with FIP200‐positive structures." k.de_vos@sheffield.ac.uk Accepted 2018-JUN-19 AT 13:01 BST AT 13:01 BST by PPORRAS The EMBO journal 2016 Only protein-protein interactions imex curation imex curation added figure 6 - two source-statements; added- a longer comment for Figs 3D/E; The other expt deleted and source text added.; added source text - lots, and altered Fig legend; added; added The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy. The EMBO journal 2016 imex curation Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. k.de_vos@sheffield.ac.uk 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ., Webster CP., Smith EF., Bauer CS., Moller A., Hautbergue GM., Ferraiuolo L., Myszczynska MA., Higginbottom A., Walsh MJ., Whitworth AJ., Kaspar BK., Meyer K., Shaw PJ., Grierson AJ., De Vos KJ. k.de_vos@sheffield.ac.uk Accepted 2018-JUN-19 AT 13:01 BST AT 13:01 BST by PPORRAS human brain random cDNA library. The EMBO journal 2016 Only protein-protein interactions imex curation imex curation added figure 6 - two source-statements; added- a longer comment for Figs 3D/E; The other expt deleted and source text added.; added source text - lots, and altered Fig legend; added; added q96lt7-2 Guanine nucleotide exchange C9orf72 C9(SF) C9-S C9orf72 protein protein human Homo sapiens Human MSTLCPPPSPAVAKTEIALSGKSPLLAATFAYWDNILGPRVRHIWAPKTEQVLLSDGEITFLANHTLNGEILRNAESGAIDVKFFVLSEKGVIIVSLIFDGNWNGDRSTYGLSIILPQTELSFYLPLHRVCVDRLTHIIRKGRIWMHKERQENVQKIILEGTERMEDQGQSIIPMLTGEVIPVMELLSSMKSHSVPEEIDIADTVLNDDDIGDSCHEGFLLK 068FE35DB48FE903 rbcc1_human RB1-inducible coiled-coil protein 1 RB1CC1 KIAA0203 RBICC FAK family kinase-interacting protein of 200 kDa protein protein human Homo sapiens Human MKLYVFLVNTGTTLTFDTELTVQTVADLKHAIQSKYKIAIQHQVLVVNGGECMAADRRVCTYSAGTDTNPIFLFNKEMILCDRPPAIPKTTFSTENDMEIKVEESLMMPAVFHTVASRTQLALEMYEVAKKLCSFCEGLVHDEHLQHQGWAAIMANLEDCSNSYQKLLFKFESIYSNYLQSIEDIKLKLTHLGTAVSVMAKIPLLECLTRHSYRECLGRLDSLPEHEDSEKAEMKRSTELVLSPDMPRTTNESLLTSFPKSVEHVSPDTADAESGKEIRESCQSTVHQQDETTIDTKDGDLPFFNVSLLDWINVQDRPNDVESLVRKCFDSMSRLDPRIIRPFIAECRQTIAKLDNQNMKAIKGLEDRLYALDQMIASCGRLVNEQKELAQGFLANQKRAENLKDASVLPDLCLSHANQLMIMLQNHRKLLDIKQKCTTAKQELANNLHVRLKWCCFVMLHADQDGEKLQALLRLVIELLERVKIVEALSTVPQMYCLAVVEVVRRKMFIKHYREWAGALVKDGKRLYEAEKSKRESFGKLFRKSFLRNRLFRGLDSWPPSFCTQKPRKFDCELPDISLKDLQFLQSFCPSEVQPFLRVPLLCDFEPLHQHVLALHNLVKAAQSLDEMSQTITDLLSEQKASVSQTSPQSASSPRMESTAGITTTTSPRTPPPLTVQDPLCPAVCPLEELSPDSIDAHTFDFETIPHPNIEQTIHQVSLDLDSLAESPESDFMSAVNEFVIEENLSSPNPISDPQSPEMMVESLYSSVINAIDSRRMQDTNVCGKEDFGDHTSLNVQLERCRVVAQDSHFSIQTIKEDLCHFRTFVQKEQCDFSNSLKCTAVEIRNIIEKVKCSLEITLKEKHQKELLSLKNEYEGKLDGLIKETEENENKIKKLKGELVCLEEVLQNKDNEFALVKHEKEAVICLQNEKDQKLLEMENIMHSQNCEIKELKQSREIVLEDLKKLHVENDEKLQLLRAELQSLEQSHLKELEDTLQVRHIQEFEKVMTDHRVSLEELKKENQQIINQIQESHAEIIQEKEKQLQELKLKVSDLSDTRCKLEVELALKEAETDEIKILLEESRAQQKETLKSLLEQETENLRTEISKLNQKIQDNNENYQVGLAELRTLMTIEKDQCISELISRHEEESNILKAELNKVTSLHNQAFEIEKNLKEQIIELQSKLDSELSALERQKDEKITQQEEKYEAIIQNLEKDRQKLVSSQEQDREQLIQKLNCEKDEAIQTALKEFKLEREVVEKELLEKVKHLENQIAKSPAIDSTRGDSSSLVAELQEKLQEEKAKFLEQLEEQEKRKNEEMQNVRTSLIAEQQTNFNTVLTREKMRKENIINDLSDKLKSTMQQQERDKDLIESLSEDRARLLEEKKKLEEEVSKLRSSSFVPSPYVATAPELYGACAPELPGESDRSAVETADEGRVDSAMETSMMSVQENIHMLSEEKQRIMLLERTLQLKEEENKRLNQRLMSQSMSSVSSRHSEKIAIRDFQVGDLVLIILDERHDNYVLFTVSPTLYFLHSESLPALDLKPGEGASGASRRPWVLGKVMEKEYCQAKKAQNRFKVPLGTKFYRVKAVSWNKKV DIP protein Q8TDY2 original sequence version: 85 DIP protein Q8TDY2 original sequence version: 95 DIP protein Q8TDY2 original sequence version: 98 DIP protein Q8TDY2 original sequence version: 107 C9C90A328875016A atg13_human Autophagy-related protein 13 ATG13 KIAA0652 protein protein human Homo sapiens Human METDLNSQDRKDLDKFIKFFALKTVQVIVQARLGEKICTRSSSSPTGSDWFNLAIKDIPEVTHEAKKALAGQLPAVGRSMCVEISLKTSEGDSMELEIWCLEMNEKCDKEIKVSYTVYNRLSLLLKSLLAITRVTPAYRLSRKQGHEYVILYRIYFGEVQLSGLGEGFQTVRVGTVGTPVGTITLSCAYRINLAFMSTRQFERTPPIMGIIIDHFVDRPYPSSSPMHPCNYRTAGEDTGVIYPSVEDSQEVCTTSFSTSPPSQLSSSRLSYQPAALGVGSADLAYPVVFAAGLNATHPHQLMVPGKEGGVPLAPNQPVHGTQADQERLATCTPSDRTHCAATPSSSEDTETVSNSSEGRASPHDVLETIFVRKVGAFVNKPINQVTLTSLDIPFAMFAPKNLELEDTDPMVNPPDSPETESPLQGSLHSDGSSGGSSGNTHDDFVMIDFKPAFSKDDILPMDLGTFYREFQNPPQLSSLSIDIGAQSMAEDLDSLPEKLAVHEKNVREFDAFVETLQ DIP protein O75143 original sequence version: 96 DIP protein O75143-1 original sequence version: 104 DIP protein O75143 original sequence version: 109 DIP protein O75143-1 original sequence version: 121 DIP protein O75143 original sequence version: 107 DIP protein O75143 original sequence version: 115 4D6905985EAB78A7 rab1a_human Ras-related protein Rab-1A YPT1-related protein RAB1A RAB1 protein protein human Homo sapiens Human MSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNVKQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC B2A8F4E3B0FB17D6 q96lt7-1 Guanine nucleotide exchange C9orf72 C9(LF) C9-L C9orf72 protein protein human Homo sapiens Human MSTLCPPPSPAVAKTEIALSGKSPLLAATFAYWDNILGPRVRHIWAPKTEQVLLSDGEITFLANHTLNGEILRNAESGAIDVKFFVLSEKGVIIVSLIFDGNWNGDRSTYGLSIILPQTELSFYLPLHRVCVDRLTHIIRKGRIWMHKERQENVQKIILEGTERMEDQGQSIIPMLTGEVIPVMELLSSMKSHSVPEEIDIADTVLNDDDIGDSCHEGFLLNAISSHLQTCGCSVVVGSSAEKVNKIVRTLCLFLTPAERKCSRLCEAESSFKYESGLFVQGLLKDSTGSFVLPFRQVMYAPYPTTHIDVDVNTVKQMPPCHEHIYNQRRYMRSELTAFWRATSEEDMAQDTIIYTDESFTPDLNIFQDVLHRDTLVKAFLDQVFQLKPGLSLRSTFLAQFLLVLHRKALTLIKYIEDDTQKGKKPFKSLRNLKIDLDLTAEGDLNIIMALAEKIKPGLHSFIFGRPFYTSVQERDVLMTF Encoded by 2 transcripts differing in the 5' non-coding region. B36C5576B3D268CE ulk1_human Serine/threonine-protein kinase ULK1 ULK1 KIAA0722 Unc-51-like kinase 1 Autophagy-related protein 1 homolog protein protein human Homo sapiens Human MEPGRGGTETVGKFEFSRKDLIGHGAFAVVFKGRHREKHDLEVAVKCINKKNLAKSQTLLGKEIKILKELKHENIVALYDFQEMANSVYLVMEYCNGGDLADYLHAMRTLSEDTIRLFLQQIAGAMRLLHSKGIIHRDLKPQNILLSNPAGRRANPNSIRVKIADFGFARYLQSNMMAATLCGSPMYMAPEVIMSQHYDGKADLWSIGTIVYQCLTGKAPFQASSPQDLRLFYEKNKTLVPTIPRETSAPLRQLLLALLQRNHKDRMDFDEFFHHPFLDASPSVRKSPPVPVPSYPSSGSGSSSSSSSTSHLASPPSLGEMQQLQKTLASPADTAGFLHSSRDSGGSKDSSCDTDDFVMVPAQFPGDLVAEAPSAKPPPDSLMCSGSSLVASAGLESHGRTPSPSPPCSSSPSPSGRAGPFSSSRCGASVPIPVPTQVQNYQRIERNLQSPTQFQTPRSSAIRRSGSTSPLGFARASPSPPAHAEHGGVLARKMSLGGGRPYTPSPQVGTIPERPGWSGTPSPQGAEMRGGRSPRPGSSAPEHSPRTSGLGCRLHSAPNLSDLHVVRPKLPKPPTDPLGAVFSPPQASPPQPSHGLQSCRNLRGSPKLPDFLQRNPLPPILGSPTKAVPSFDFPKTPSSQNLLALLARQGVVMTPPRNRTLPDLSEVGPFHGQPLGPGLRPGEDPKGPFGRSFSTSRLTDLLLKAAFGTQAPDPGSTESLQEKPMEIAPSAGFGGSLHPGARAGGTSSPSPVVFTVGSPPSGSTPPQGPRTRMFSAGPTGSASSSARHLVPGPCSEAPAPELPAPGHGCSFADPITANLEGAVTFEAPDLPEETLMEQEHTEILRGLRFTLLFVQHVLEIAALKGSASEAAGGPEYQLQESVVADQISLLSREWGFAEQLVLYLKVAELLSSGLQSAIDQIRAGKLCLSSTVKQVVRRLNELYKASVVSCQGLSLRLQRFFLDKQRLLDRIHSITAERLIFSHAVQMVQSAALDEMFQHREGCVPRYHKALLLLEGLQHMLSDQADIENVTKCKLCIERRLSALLTGICA DIP protein O75385 original sequence version: 129 DIP protein O75385 original sequence version: 141 DIP protein O75385 original sequence version: 133 DIP protein O75385 original sequence version: 132 DIP protein O75385 original sequence version: 142 AED9BD5139F2DB92 ci072_human Guanine nucleotide exchange C9orf72 C9orf72 protein protein human Homo sapiens Human MSTLCPPPSPAVAKTEIALSGKSPLLAATFAYWDNILGPRVRHIWAPKTEQVLLSDGEITFLANHTLNGEILRNAESGAIDVKFFVLSEKGVIIVSLIFDGNWNGDRSTYGLSIILPQTELSFYLPLHRVCVDRLTHIIRKGRIWMHKERQENVQKIILEGTERMEDQGQSIIPMLTGEVIPVMELLSSMKSHSVPEEIDIADTVLNDDDIGDSCHEGFLLNAISSHLQTCGCSVVVGSSAEKVNKIVRTLCLFLTPAERKCSRLCEAESSFKYESGLFVQGLLKDSTGSFVLPFRQVMYAPYPTTHIDVDVNTVKQMPPCHEHIYNQRRYMRSELTAFWRATSEEDMAQDTIIYTDESFTPDLNIFQDVLHRDTLVKAFLDQVFQLKPGLSLRSTFLAQFLLVLHRKALTLIKYIEDDTQKGKKPFKSLRNLKIDLDLTAEGDLNIIMALAEKIKPGLHSFIFGRPFYTSVQERDVLMTF B36C5576B3D268CE c9orf72-1 5 13 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 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 physical association physical association Figure 3 A c9orf72-rb1cc1-1 5 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 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 physical association physical association Figure 3 A-2 FLAG‐FIP200, HA‐ULK1, and Myc‐ATG13 specifically co‐immunoprecipitated with C9orf72S and C9orf72L. c9orf72-rb1cc1-2 5 13 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 14 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed n_terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 3B FLAG‐FIP200, HA‐ULK1, and Myc‐ATG13 specifically co‐immunoprecipitated with C9orf72S and C9orf72L. 2345etr-1 5 11 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 13 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 physical association physical association Fig 3C c9orf72-atg13-1 5 11 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 physical association physical association Fig 3C-2 c9orf72-rb1cc1-3 3 11 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 13 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role prey prey association association Figure 3 D we immunoprecipitated transfected Myc‐tagged C9orf72S or C9orf72L and probed the resulting immune pellet for endogenous FIP200, ULK1, and ATG13. All three endogenous members of the ULK1 initiation complex were detected in the C9orf72 immunoprecipitates c9orf72-atg13-2 3 11 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role prey prey association association Figure 3 D we immunoprecipitated transfected Myc‐tagged C9orf72S or C9orf72L and probed the resulting immune pellet for endogenous FIP200, ULK1, and ATG13. All three endogenous members of the ULK1 initiation complex were detected in the C9orf72 immunoprecipitates c9orf72-ulk1-1 5 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 14 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed n_terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 3B-2 FLAG‐FIP200, HA‐ULK1, and Myc‐ATG13 specifically co‐immunoprecipitated with C9orf72S and C9orf72L. c9orf72-atg13-3 3 14 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 3 E we immunoprecipitated transfected Myc‐tagged C9orf72S or C9orf72L and probed the resulting immune pellet for endogenous FIP200, ULK1, and ATG13. All three endogenous members of the ULK1 initiation complex were detected in the C9orf72 immunoprecipitates c9orf72-atg13-4 3 14 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 13 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 3 E we immunoprecipitated transfected Myc‐tagged C9orf72S or C9orf72L and probed the resulting immune pellet for endogenous FIP200, ULK1, and ATG13. All three endogenous members of the ULK1 initiation complex were detected in the C9orf72 immunoprecipitates 436453-1 2 13 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 14 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n_terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range proximity proximity close-contact colocalization neighbourhood interaction Fig 3G, EV2 Using antibodies against the tags, we observed proximity signals in all co‐transfected cells examined (Figs 3F–H and EV2), confirming the interaction of C9orf72 with the ULK1 initiation complex in intact cells c9orf72-ulk1-2 2 9 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 14 unspecified role unspecified role neutral component neutral component n_terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range proximity proximity close-contact colocalization neighbourhood interaction Fig 3 G-2, EV2 Using antibodies against the tags, we observed proximity signals in all co‐transfected cells examined (Figs 3F–H and EV2), confirming the interaction of C9orf72 with the ULK1 initiation complex in intact cells c9orf72-ulk1-3 2 13 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed 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 proximity proximity close-contact colocalization neighbourhood interaction Fig 3F, EV2 Using antibodies against the tags, we observed proximity signals in all co‐transfected cells examined (Figs 3F–H and EV2), confirming the interaction of C9orf72 with the ULK1 initiation complex in intact cells c9orf72-rb1cc1-4 2 9 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed 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 proximity proximity close-contact colocalization neighbourhood interaction Fig 3 F-2, EV2 Using antibodies against the tags, we observed proximity signals in all co‐transfected cells examined (Figs 3F–H and EV2), confirming the interaction of C9orf72 with the ULK1 initiation complex in intact cells c9orf72-rb1cc1-5 2 9 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n_terminus egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 11 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range proximity proximity close-contact colocalization neighbourhood interaction Fig 3 H-2, EV2 Using antibodies against the tags, we observed proximity signals in all co‐transfected cells examined (Figs 3F–H and EV2), confirming the interaction of C9orf72 with the ULK1 initiation complex in intact cells atg13-c9orf72-2 2 11 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 13 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n_terminus egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range proximity proximity close-contact colocalization neighbourhood interaction Fig 3H, EV2 Using antibodies against the tags, we observed proximity signals in all co‐transfected cells examined (Figs 3F–H and EV2), confirming the interaction of C9orf72 with the ULK1 initiation complex in intact cells 2345rr-1 4 13 unspecified role unspecified role bait bait purified purified n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role prey prey cell lysate cell lysate region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n_terminal_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro physical association physical association 3I C9orf72S and C9orf72L bound to the N‐terminus of FIP200 (aa 1–638), but also bound non-specifically to the C-terminal region of FIP200. C9orf72S and C9orf72L bound to the N‐terminus of FIP200 (aa 1–638) c9orf72-rb1cc1-6 4 9 unspecified role unspecified role bait bait purified purified n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role prey prey cell lysate cell lysate region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n_terminal_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro physical association physical association 3Ii c9orf72-rb1cc1-7 4 13 unspecified role unspecified role bait bait purified purified n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role prey prey cell lysate cell lysate n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association 3J C9orf72S and C9orf72L also interacted with ATG13 and ULK1 (Fig 3J and K). c9orf72-atg13-5 4 9 unspecified role unspecified role bait bait purified purified n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role prey prey cell lysate cell lysate n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association 3J-2 C9orf72S and C9orf72L also interacted with ATG13 and ULK1 (Fig 3J and K). c9orf72-atg13-6 4 13 unspecified role unspecified role bait bait purified purified n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 14 unspecified role unspecified role prey prey cell lysate cell lysate n_terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association 3K C9orf72S and C9orf72L also interacted with ATG13 and ULK1 (Fig 3J and K). c9orf72-ulk1-4 4 9 unspecified role unspecified role bait bait purified purified n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 14 unspecified role unspecified role prey prey cell lysate cell lysate region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n_terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range in vitro In vitro physical association physical association 3K-2 C9orf72S and C9orf72L also interacted with ATG13 and ULK1 (Fig 3J and K). 23142q3eqrq-1 1 10 tag fluorescence tag visualisation by fluorescence tag fluorescence immunocytochemistry immunocytochemistry unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell n_terminus mcherry mcherry fluorescent protein tag mcherry n-term range n-terminal range n-term range n-term range n-terminal range n-term range 13 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range colocalization colocalization 5B Torin1- an inhibitor of mTOR and inducer of ULK1‐dependent autophagy. Rab1a siRNA c9orf72-rb1cc1-8 7 9 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 tag fluorescence tag visualisation by fluorescence tag fluorescence immunocytochemistry immunocytochemistry unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell n_terminus mcherry mcherry fluorescent protein tag mcherry n-term range n-terminal range n-term range n-term range n-terminal range n-term range colocalization colocalization 4D, 5A, EV3 Rab1a siRNA Torin1- an inhibitor of mTOR and inducer of ULK1‐dependent autophagy. c9orf72-rb1cc1-9 1 9 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 tag fluorescence tag visualisation by fluorescence tag fluorescence immunocytochemistry immunocytochemistry unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell n_terminus mcherry mcherry fluorescent protein tag mcherry n-term range n-terminal range n-term range n-term range n-terminal range n-term range colocalization colocalization 5B Torin1- an inhibitor of mTOR and inducer of ULK1‐dependent autophagy. rab1A siRNA c9orf72-rb1cc1-10 7 10 tag fluorescence tag visualisation by fluorescence tag fluorescence immunocytochemistry immunocytochemistry unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell n_terminus mcherry mcherry fluorescent protein tag mcherry n-term range n-terminal range n-term range n-term range n-terminal range n-term range 13 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range colocalization colocalization 4D, 5A, EV3 Torin1- an inhibitor of mTOR and inducer of ULK1‐dependent autophagy. Rab1a siRNA 678yuiiop-1 8 12 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association Fig 6A Consistent with a direct interaction, we isolated a partial Rab1a clone (aa 46–205) in a Y2H screen of a human brain cDNA library with C9orf72S as “bait” (Fig 6A). c9orf72-atg13-7 5 12 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 13 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 physical association physical association Fig 6B Rab1a co‐immunoprecipitated efficiently with C9orf72L and to a lesser extent with C9orf72S c9orf72-rab1a-1 5 12 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 physical association physical association Fig 6B-2 Rab1a co‐immunoprecipitated efficiently with C9orf72L and to a lesser extent with C9orf72S c9orf72-rab1a-2 3 13 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 12 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Fig 6C Rab1a co‐immunoprecipitated efficiently with C9orf72L and to a lesser extent with C9orf72S c9orf72-rab1a-3 3 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 12 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Fig 6C-2 Rab1a co‐immunoprecipitated efficiently with C9orf72L and to a lesser extent with C9orf72S c9orf72-atg13-8 4 9 unspecified role unspecified role bait bait purified purified n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role prey prey cell lysate cell lysate region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range in vitro In vitro physical association physical association 6D non‐hydrolysable GTP analog GMP‐PNP (CHEBI:78408) Rab1a interacted with both C9orf72L and C9orf72S in these assays (Fig 6D) c9orf72-rab1a-4 4 13 unspecified role unspecified role bait bait purified purified n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role prey prey cell lysate cell lysate region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range in vitro In vitro physical association physical association 6D-2 non‐hydrolysable GTP analog GMP‐PNP (CHEBI:78408) Rab1a interacted with both C9orf72L and C9orf72S c9orf72-rab1a-5 4 13 unspecified role unspecified role prey prey cell lysate cell lysate region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range in vitro In vitro 12 unspecified role unspecified role bait bait purified purified region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 6E C9orf72 binds to Rab1a and behaves as a Rab1a effector rather than a Rab1a GEF. The binding curve fitted a hyperbola (R2 = 0.94). C9orf72 binds to Rab1a and behaves as a Rab1a effector rather than a Rab1a GEF. rab1a-ulk1-1 6 15 unspecified role unspecified role ancillary ancillary physiological level physiological level endogenous level endogenous endogenous endogenous level 12 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 14 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n_terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range proximity proximity close-contact colocalization neighbourhood interaction Fig 7A-RHS image we observed proximity signals in all cells co‐transfected with HA‐ULK1 and Myc‐Rab1a while in cells transfected with HA‐ULK1 or Myc‐Rab1a alone only very low numbers of proximity signals were observed. these data show ULK1–Rab1a interaction in intact cells and reveal that this interaction is C9orf72 dependent c9orf72-rb1cc1-11 7 12 tag fluorescence tag visualisation by fluorescence tag fluorescence immunocytochemistry immunocytochemistry unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell n_terminus mcherry mcherry fluorescent protein tag mcherry n-term range n-terminal range n-term range n-term range n-terminal range n-term range P62820:p.Gln70Leu mutation mutation certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 16:49:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln70leu' Tue Jun 30 19:55:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln70Leu' 13 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range colocalization colocalization 7B C9orf72 siRNA I can still some co-loc in presence of C9orf72 siRNA (orange colour)