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/ Proteasome-mediated turnover of the transcription coactivator NPR1 plays dual roles in regulating plant immunity. Proteasome-mediated turnover of the transcription coactivator NPR1 plays dual roles in regulating plant immunity. Cell (0092-8674) 2009 imex curation Spoel SH., Mou Z., Tada Y., Spivey NW., Genschik P., Dong X. Experiments performed in transgenic 35S::NPR1-GFP plants. An interaction was also observed between NPR1 and CSN5 but this could not be ascribed to a single gene product. xdong@duke.edu davet Only protein-protein interactions 5 in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Spoel SH., Mou Z., Tada Y., Spivey NW., Genschik P., Dong X. Experiments performed in transgenic 35S::NPR1-GFP plants. An interaction was also observed between NPR1 and CSN5 but this could not be ascribed to a single gene product. xdong@duke.edu Accepted 2009-AUG-16 by DAVET. Cell (0092-8674) 2009 davet Only protein-protein interactions 5 imex curation Proteasome-mediated turnover of the transcription coactivator NPR1 plays dual roles in regulating plant immunity. Proteasome-mediated turnover of the transcription coactivator NPR1 plays dual roles in regulating plant immunity. Cell (0092-8674) 2009 imex curation Spoel SH., Mou Z., Tada Y., Spivey NW., Genschik P., Dong X. Experiments performed in transgenic 35S::NPR1-GFP plants. An interaction was also observed between NPR1 and CSN5 but this could not be ascribed to a single gene product. xdong@duke.edu davet Only protein-protein interactions 5 arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Spoel SH., Mou Z., Tada Y., Spivey NW., Genschik P., Dong X. Experiments performed in transgenic 35S::NPR1-GFP plants. An interaction was also observed between NPR1 and CSN5 but this could not be ascribed to a single gene product. xdong@duke.edu Accepted 2009-AUG-16 by DAVET. Cell (0092-8674) 2009 davet Only protein-protein interactions 5 imex curation csn4_arath COP9 signalosome complex subunit 4 Constitutive photomorphogenesis protein 8 Protein FUSCA 4 CSN4 COP8 FUS4 At5g42970 MBD2.17 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDEALTNASAIGDQRQKIEQYKLILSSVLSSNDLLQAQRFIDHILSDDVPLVVSRQLLQSFAQELGRLEPETQKEIAQFTLTQIQPRVVSFEEQALVIREKLAGLYESEQEWSKAAQMLSGIDLDSGMRAVDDNFKLSKCIQIARLYLEDDDAVNAEAFINKASFLVSNSQNEVLNLQYKVCYARILDMKRKFLEAALRYYGISQIEQRQIGDEEIDENALEQALSAAVTCTILAGAGPQRSRVLATLYKDERCSKLKIYPILQKVYLERILRRPEIDAFSEELRPHQKASLPDKSTVLDRAMIEHNLLSASKLYTNIRFDELGTLLAIDPRKAEKIAANMIGQDRMRGSIDQEEAVIHFEDDVEELQQWDQQISGLCQALNDILDGMAKKGMSVPV 1E8AEF28C35B28F0 npr1_arath Regulatory protein NPR1 SAI1 BTB/POZ domain-containing protein NPR1 NPR1 NIM1 At1g64280 F15H21.6 Nonexpresser of PR genes 1 Non-inducible immunity protein 1 Salicylic acid insensitive 1 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDTTIDGFADSYEISSTSFVATDNTDSSIVYLAAEQVLTGPDVSALQLLSNSFESVFDSPDDFYSDAKLVLSDGREVSFHRCVLSARSSFFKSALAAAKKEKDSNNTAAVKLELKEIAKDYEVGFDSVVTVLAYVYSSRVRPPPKGVSECADENCCHVACRPAVDFMLEVLYLAFIFKIPELITLYQRHLLDVVDKVVIEDTLVILKLANICGKACMKLLDRCKEIIVKSNVDMVSLEKSLPEELVKEIIDRRKELGLEVPKVKKHVSNVHKALDSDDIELVKLLLKEDHTNLDDACALHFAVAYCNVKTATDLLKLDLADVNHRNPRGYTVLHVAAMRKEPQLILSLLEKGASASEATLEGRTALMIAKQATMAVECNNIPEQCKHSLKGRLCVEILEQEDKREQIPRDVPPSFAVAADELKMTLLDLENRVALAQRLFPTEAQAAMEIAEMKGTCEFIVTSLEPDRLTGTKRTSPGVKIAPFRILEEHQSRLKALSKTVELGKRFFPRCSAVLDQIMNCEDLTQLACGEDDTAEKRLQKKQRYMEIQETLKKAFSEDNLELGNSSLTDSTSSTSKSTGGKRSNRKLSHRRR DIP protein P93002 original sequence version: 106 DDD618146254CD68 csn8_arath COP9 signalosome complex subunit 8 Constitutive photomorphogenesis protein 9 Protein FUSCA 7 CSN8 COP9 FUS7 At4g14110 dl3095c protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDLSPVKEALAAKSFDKIADICDTLMLQVASEGIEYHDDWPYAIHLLGYFYVDDCDSARFLWKRIPTAIKERKPEVVAAWGIGQKLWTHDYAGVYEAIRGYDWSQEAKDMVAAFSDLYTKRMFQLLLSAYSTITIHDLALFLGMTEDDATTYVVENGWTVDAASQMASVKKQAVKREQKVDSSKLQRLTEYVFHLEH D182ABACE4238759 tga2_arath Transcription factor TGA2 TGA2 BZIP20 HBP1B At5g06950 MOJ9.12 HBP-1b homolog bZIP transcription factor 20 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MADTSPRTDVSTDDDTDHPDLGSEGALVNTAASDSSDRSKGKMDQKTLRRLAQNREAARKSRLRKKAYVQQLENSRLKLTQLEQELQRARQQGVFISGTGDQAHSTGGNGALAFDAEHSRWLEEKNKQMNELRSALNAHAGDSELRIIVDGVMAHYEELFRIKSNAAKNDVFHLLSGMWKTPAERCFLWLGGFRSSELLKLLANQLEPMTERQLMGINNLQQTSQQAEDALSQGMESLQQSLADTLSSGTLGSSSSGNVASYMGQMAMAMGKLGTLEGFIRQADNLRLQTLQQMIRVLTTRQSARALLAIHDYFSRLRALSSLWLARPRE D3033153BCE932C4 cul3a_arath Cullin-3A CUL3A T2P11.2 CUL3 At1g26830 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSNQKKRNFQIEAFKHRVVVDPKYADKTWQILERAIHQIYNQDASGLSFEELYRNAYNMVLHKFGEKLYTGFIATMTSHLKEKSKLIEAAQGGSFLEELNKKWNEHNKALEMIRDILMYMDRTYIESTKKTHVHPMGLNLWRDNVVHFTKIHTRLLNTLLDLVQKERIGEVIDRGLMRNVIKMFMDLGESVYQEDFEKPFLDASSEFYKVESQEFIESCDCGDYLKKSEKRLTEEIERVAHYLDAKSEEKITSVVEKEMIANHMQRLVHMENSGLVNMLLNDKYEDLGRMYNLFRRVTNGLVTVRDVMTSHLREMGKQLVTDPEKSKDPVEFVQRLLDERDKYDKIINTAFGNDKTFQNALNSSFEYFINLNARSPEFISLFVDDKLRKGLKGITDVDVEVILDKVMMLFRYLQEKDVFEKYYKQHLAKRLLSGKTVSDDAERSLIVKLKTECGYQFTSKLEGMFTDMKTSEDTMRGFYGSHPELSEGPTLIVQVLTTGSWPTQPAVPCNLPAEVSVLCEKFRSYYLGTHTGRRLSWQTNMGTADIKAIFGKGQKHELNVSTFQMCVLMLFNNSDRLSYKEIEQATEIPAADLKRCLQSLACVKGKNVIKKEPMSKDIGEEDLFVVNDKFTSKFYKVKIGTVVAQKETEPEKQETRQRVEEDRKPQIEAAIVRIMKSRKILDHNNIIAEVTKQLQPRFLANPTEIKKRIESLIERDFLERDSTDRKLYRYLA DIP protein Q9ZVH4 original sequence version: 81 40F58A46BB5572ED npr1-cul3a-1 2 7 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Total and immunoprecipitated proteins were analyzed by western blotting using anti-GFP and anti-CUL3A antibodies. Figs. 2A and 3A Salicylic acid npr1-cop9-1 2 4 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role bait bait physical association physical association Unexpectedly, coimmunoprecipitation experiments indicated that in SA treated plants, more NPR1-GFP was pulled down with CUL3A and three components of the CSN complex (COP9, CSN4, CSN5; Figure 3A). Fig. 3A Salicylic acid npr1-csn4-1 2 4 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 3 unspecified role unspecified role bait bait physical association physical association Unexpectedly, coimmunoprecipitation experiments indicated that in SA treated plants, more NPR1-GFP was pulled down with CUL3A and three components of the CSN complex (COP9, CSN4, CSN5; Figure 3A). Fig. 3A Salicylic acid npr1-cul3a-2 2 4 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P93002:p.[Ser11Ala;Ser15Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S A certain certain sequence position certain certain certain sequence position certain S A Wed May 02 19:46:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser11ala,ser15ala' Tue Jun 30 18:11:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser11Ala;Ser15Ala]' 7 unspecified role unspecified role bait bait physical association physical association Total and immunoprecipitated proteins were analyzed by western blotting using anti-GFP and anti-CUL3A antibodies. Fig. 6B Salicylic acid and proteasome inhibitor MG115 Salicylic acid-induced phosphorylation of NPR1 facilitates its interaction with the CUL3-based ubiquitin ligase npr1-tga2-9 1 4 unspecified role unspecified role bait bait region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress 6 unspecified role unspecified role prey prey 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 direct interaction direct interaction The phosphorylation status of NPR1 does not affect interaction with TGA transcription factors Supp. Fig. 9 Salicylic acid