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/ Membrane association of the Arabidopsis ARF exchange factor GNOM involves interaction of conserved domains. Membrane association of the Arabidopsis ARF exchange factor GNOM involves interaction of conserved domains. Plant Cell (1040-4651) 2008 imex curation Anders N., Nielsen M., Keicher J., Stierhof YD., Furutani M., Tasaka M., Skriver K., Jurgens G. gerd.juergens@zmbp.uni-tuebingen.de. 27-Oct-2007: Contacted by JYOTI. orchard Only protein-protein interactions arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Anders N., Nielsen M., Keicher J., Stierhof YD., Furutani M., Tasaka M., Skriver K., Jurgens G. gerd.juergens@zmbp.uni-tuebingen.de. Accepted 2009-NOV-5 by ORCHARD. Plant Cell (1040-4651) 2008 orchard Only protein-protein interactions imex curation Membrane association of the Arabidopsis ARF exchange factor GNOM involves interaction of conserved domains. Membrane association of the Arabidopsis ARF exchange factor GNOM involves interaction of conserved domains. Plant Cell (1040-4651) 2008 imex curation Anders N., Nielsen M., Keicher J., Stierhof YD., Furutani M., Tasaka M., Skriver K., Jurgens G. gerd.juergens@zmbp.uni-tuebingen.de. 27-Oct-2007: Contacted by JYOTI. orchard Only protein-protein interactions 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 predetermined predetermined participant predetermined Anders N., Nielsen M., Keicher J., Stierhof YD., Furutani M., Tasaka M., Skriver K., Jurgens G. gerd.juergens@zmbp.uni-tuebingen.de. Accepted 2009-NOV-5 by ORCHARD. 27-Oct-2007: Contacted by JYOTI. Plant Cell (1040-4651) 2008 orchard Only protein-protein interactions imex curation Membrane association of the Arabidopsis ARF exchange factor GNOM involves interaction of conserved domains. Membrane association of the Arabidopsis ARF exchange factor GNOM involves interaction of conserved domains. Plant Cell (1040-4651) 2008 imex curation Anders N., Nielsen M., Keicher J., Stierhof YD., Furutani M., Tasaka M., Skriver K., Jurgens G. gerd.juergens@zmbp.uni-tuebingen.de. 27-Oct-2007: Contacted by JYOTI. orchard Only protein-protein interactions arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column anti tag western anti tag western blot anti tag western Anders N., Nielsen M., Keicher J., Stierhof YD., Furutani M., Tasaka M., Skriver K., Jurgens G. gerd.juergens@zmbp.uni-tuebingen.de. Accepted 2009-NOV-5 by ORCHARD. 27-Oct-2007: Contacted by JYOTI. Plant Cell (1040-4651) 2008 orchard Only protein-protein interactions imex curation gnom_arath ARF guanine-nucleotide exchange factor GNOM GN EMB30 GBF3 MIZ2 Pattern formation protein EMB30 Protein EMBRYO DEFECTIVE 30 Protein MIZU-KUSSEI2 Protein VASCULAR NETWORK 7 VAN7 F16A14.20 F7A19.7 At1g13980 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGRLKLHSGIKAIEEEPEDFECTDSSNTTTLACMIDTEIAAVLAVMRRNVRWGGRYMSGDDQLEHSLIQSLKALRKQVFSWNQPWHTISPMLYLQPFLDVIRSDETGAPITSIALSSVYKILNLNVIDQNTANIEDAMHLVVDSVTSCRFEVTDPASEEVVLMKILQVLLACMKNKASVMLSNQHVCTVVNTCFRVVHQAGMKGELLQRVARHTMHELVRCIFSHLPDVERTETTLVNRAGSIKQEKAGVDSDYAIVSKPVEDGNANSEYDVENSMATFATGAQSLMDDGPVGPGSRKPASPYDLHIMTEPYGVPSMVEIFHFLCSLLNVVEHVGMGSRSNTIAFDEDVPLFALNLINSAIELGGSSIRHHPRLLSLIQDELFRNLMQFGLSMSPLILSMVCSIVLNLYQHLRTELKLQLEAFFSCVILRLAQGKYGPSYQQQEVAMEALVNFCRQKSFMVEMYANLDCDITCSNVFEELSNLLSKSTFPVNCPLSAMHILALDGLIAVIQGMAERISNGLTGLDLGPVHLDEYTPFWMVKCDNYSDPNHWVSFVRRRKYIKRRLMIGADHFNRDPKKGLEFLQGTHLLPDKLDPQSVACFFRYTAGLDKNLVGDFLGNHDEFCVQVLNEFAGTFDFQYMNLDTALRLFLETFRLPGESQKIQRVLEAFSERYYMQSPEILANKDAALVLSYSIIMLNTDQHNVQVKKKMTEEDFIRNNRHINGGNDLPREFLSELFHSICNNEIRTTPEQGAGFPEMTPSRWIDLMHKSKKTAPYILADSRAYLDHDMFAIMSGPTIAAISVVFDHAEHEDVYQTCIDGFLAIAKISACHHLEDVLDDLVVSLCKFTTLLNPSSVDEPVLAFGDDAKARMATITIFTIANKYGDYIRTGWRNILDCILRLHKLGLLPARVASDAADESEHSSEQGQGKPLANSLSSAHLQSMGTPRRSSGLMGRFSQLLSLDTEEPRSQPTEQQLAAHQRTLQTIQKCHIDSIFTESKFLQAESLLQLARALIWAAGRPQKGTSSPEDEDTAVFCLELLIAITLNNRDRIVLLWQGVYEHIATIAQSTVMPCNLVDKAIFGLLRICQRLLPYKESLADELLRSLQLVLKLDARVADAYCEQIAIEVSRLVKANANHIRSQAGWRTITSLLSITARHPEASESGFDAVSFVMSEGTHLYPANYVLCVDAARQFAESRVGQSERSIRALDLMGDSLEFLAKWALSAKENMGEEDFGKMSQDIGEMWLRLVQGLRKVCLDQREDVRNHALQSLQKCLGGVDGINLAHSMWSQCFDKVIFTVLDDLLEIAAGSQKDYRNMEGTLLLAIKLLSKVFLQQLQELSQLSTFCKLWLGVLTRMEKYMKVKVRGKKSDKLQESVPELLKNILLVMKTKGVLLQRSALGGDSLWELTWLHVNNIAPSMRLELFPDQESSQLGDDETVSNGLSSPENTTGS 666E21C74B426996 gnom-gnom 1 4 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 4 unspecified role unspecified role prey prey n-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 8 physical association physical association Figure 1B GNOM-HA interacted with GNOM-Myc, whereas no in vivo interaction was observed between GNOMΔDCB variants (Figure 1B) in coimmunoprecipitation gnom-gnom-1 2 4 unspecified role unspecified role bait bait n-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 4 unspecified role unspecified role prey prey n-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Supplementary Figure 1 Figure 3A Homotypic interaction of GNOMΔDCB and the DCB domain (positive control). Note that GNOMΔDCB does not interact with itself. gnom-gnom-2 3 4 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Figure 1C Thus, the DCB�DCB domain interaction mediates the formation of GNOM dimers, rather than oligomers, as its most abundant form in vivo. gnom-gnom-3 2 4 unspecified role unspecified role bait bait n-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 4 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q42510:p.Asp468Gly mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain D G Wed May 02 20:14:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp468gly' Tue Jun 30 18:30:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp468Gly' Q42510:p.Gly579Arg mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain G R Wed May 02 14:50:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly579arg' Tue Jun 30 17:53:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly579Arg' physical association physical association Supplementary Figure 1 Figure 3A Figure 3D To assess its essential, unidentified function, the DCB domain was tested for interaction with other domains of the GNOM protein by yeast two-hybrid assay. Surprisingly, the DCB domain interacted not only with itself (homotypic interaction) but also remarkably strongly with GNOM lacking the DCB domain (GNOMΔDCB; heterotypic interaction) (Figure 3A) gnom-gnom-4 1 4 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 34 30 physical association physical association Figure 3B To determine whether the heterotypic interaction also occurs in vivo, we coexpressed fragments of GNOM protein in transgenic plants. Coimmunoprecipitation of both DCB-Myc and full-length GNOM-Myc with GNOMΔDCB-HA revealed the heterotypic interaction