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/ Growth arrest and DNA damage-inducible protein GADD34 assembles a novel signaling complex containing protein phosphatase 1 and inhibitor 1. Growth arrest and DNA damage-inducible protein GADD34 assembles a novel signaling complex containing protein phosphatase 1 and inhibitor 1. Mol. Cell. Biol. (0270-7306) 2001 imex curation Connor JH., Weiser DC., Li S., Hallenbeck JM., Shenolikar S. Apoptosis - Interactions involving proteins with a function related to apoptosis Data on rabbit PP1 lost as mammals have at least 3 gene products giving a catalytic PP1 subunit. Squirrel data also lost as NEWT lists about 200 squirrels sheno001@mc.duke.edu 01-April-2008: Contacted by JYOTI. Human brain cDNA davet 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Connor JH., Weiser DC., Li S., Hallenbeck JM., Shenolikar S. Apoptosis - Interactions involving proteins with a function related to apoptosis Data on rabbit PP1 lost as mammals have at least 3 gene products giving a catalytic PP1 subunit. Squirrel data also lost as NEWT lists about 200 squirrels sheno001@mc.duke.edu Accepted 2008-FEB-3 by DAVET. 01-April-2008: Contacted by JYOTI. Human brain cDNA Mol. Cell. Biol. (0270-7306) 2001 davet Only protein-protein interactions imex curation Growth arrest and DNA damage-inducible protein GADD34 assembles a novel signaling complex containing protein phosphatase 1 and inhibitor 1. Growth arrest and DNA damage-inducible protein GADD34 assembles a novel signaling complex containing protein phosphatase 1 and inhibitor 1. Mol. Cell. Biol. (0270-7306) 2001 imex curation Connor JH., Weiser DC., Li S., Hallenbeck JM., Shenolikar S. Apoptosis - Interactions involving proteins with a function related to apoptosis Data on rabbit PP1 lost as mammals have at least 3 gene products giving a catalytic PP1 subunit. Squirrel data also lost as NEWT lists about 200 squirrels sheno001@mc.duke.edu 01-April-2008: Contacted by JYOTI. Human brain cDNA davet 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 Connor JH., Weiser DC., Li S., Hallenbeck JM., Shenolikar S. Apoptosis - Interactions involving proteins with a function related to apoptosis Data on rabbit PP1 lost as mammals have at least 3 gene products giving a catalytic PP1 subunit. Squirrel data also lost as NEWT lists about 200 squirrels sheno001@mc.duke.edu Accepted 2008-FEB-3 by DAVET. 01-April-2008: Contacted by JYOTI. Human brain cDNA Mol. Cell. Biol. (0270-7306) 2001 davet Only protein-protein interactions imex curation Growth arrest and DNA damage-inducible protein GADD34 assembles a novel signaling complex containing protein phosphatase 1 and inhibitor 1. Growth arrest and DNA damage-inducible protein GADD34 assembles a novel signaling complex containing protein phosphatase 1 and inhibitor 1. Mol. Cell. Biol. (0270-7306) 2001 imex curation Connor JH., Weiser DC., Li S., Hallenbeck JM., Shenolikar S. Apoptosis - Interactions involving proteins with a function related to apoptosis Data on rabbit PP1 lost as mammals have at least 3 gene products giving a catalytic PP1 subunit. Squirrel data also lost as NEWT lists about 200 squirrels sheno001@mc.duke.edu 01-April-2008: Contacted by JYOTI. Human brain cDNA davet Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Connor JH., Weiser DC., Li S., Hallenbeck JM., Shenolikar S. Apoptosis - Interactions involving proteins with a function related to apoptosis Data on rabbit PP1 lost as mammals have at least 3 gene products giving a catalytic PP1 subunit. Squirrel data also lost as NEWT lists about 200 squirrels I-1 was coupled to CNBr-activated Sepharose sheno001@mc.duke.edu Accepted 2008-FEB-3 by DAVET. 01-April-2008: Contacted by JYOTI. Human brain cDNA Mol. Cell. Biol. (0270-7306) 2001 davet Only protein-protein interactions imex curation pp12_yeast Serine/threonine-protein phosphatase PP1-2 GLC7 CID1 DIS2 YER133W protein protein yeast Saccharomyces cerevisiae Baker's yeast MDSQPVDVDNIIDRLLEVRGSKPGQQVDLEENEIRYLCSKARSIFIKQPILLELEAPIKICGDIHGQYYDLLRLFEYGGFPPESNYLFLGDYVDRGKQSLETICLLLAYKIKYPENFFILRGNHECASINRIYGFYDECKRRYNIKLWKTFTDCFNCLPIAAIIDEKIFCMHGGLSPDLNSMEQIRRVMRPTDIPDVGLLCDLLWSDPDKDIVGWSENDRGVSFTFGPDVVNRFLQKQDMELICRAHQVVEDGYEFFSKRQLVTLFSAPNYCGEFDNAGAMMSVDESLLCSFQILKPAQKSLPRQAGGRKKK 71B20E3A4ED93235 71B20E3A4ED93235 ppr1a_human Protein phosphatase 1 regulatory subunit 1A Protein phosphatase inhibitor 1 PPP1R1A IPP1 protein protein human Homo sapiens Human MEQDNSPRKIQFTVPLLEPHLDPEAAEQIRRRRPTPATLVLTSDQSSPEIDEDRIPNPHLKSTLAMSPRQRKKMTRITPTMKELQMMVEHHLGQQQQGEEPEGAAESTETQESRPPGIPDTEVESRLGTSGTAKKTAECIPKTHERGSKEPSTKEPSTHIPPLDSKGANSV 6F61FF08E4305729 pr15a_human Protein phosphatase 1 regulatory subunit 15A Myeloid differentiation primary response protein MyD116 homolog PPP1R15A GADD34 Growth arrest and DNA damage-inducible protein GADD34 protein protein human Homo sapiens Human MAPGQAPHQATPWRDAHPFFLLSPVMGLLSRAWSRLRGLGPLEPWLVEAVKGAALVEAGLEGEARTPLAIPHTPWGRRPEEEAEDSGGPGEDRETLGLKTSSSLPEAWGLLDDDDGMYGEREATSVPRGQGSQFADGQRAPLSPSLLIRTLQGSDKNPGEEKAEEEGVAEEEGVNKFSYPPSHRECCPAVEEEDDEEAVKKEAHRTSTSALSPGSKPSTWVSCPGEEENQATEDKRTERSKGARKTSVSPRSSGSDPRSWEYRSGEASEEKEEKAHKETGKGEAAPGPQSSAPAQRPQLKSWWCQPSDEEEGEVKALGAAEKDGEAECPPCIPPPSAFLKAWVYWPGEDTEEEEDEEEDEDSDSGSDEEEGEAEASSSTPATGVFLKSWVYQPGEDTEEEEDEDSDTGSAEDEREAETSASTPPASAFLKAWVYRPGEDTEEEEDEDVDSEDKEDDSEAALGEAESDPHPSHPDQRAHFRGWGYRPGKETEEEEAAEDWGEAEPCPFRVAIYVPGEKPPPPWAPPRLPLRLQRRLKRPETPTHDPDPETPLKARKVRFSEKVTVHFLAVWAGPAQAARQGPWEQLARDRSRFARRITQAQEELSPCLTPAARARAWARLRNPPLAPIPALTQTLPSSSVPSSPVQTTPLSQAVATPSRSSAAAAAALDLSGRRG B257AA17456D1403 microcystin-lr Microcystin LR small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) ppp1r1a-ppp1r15a-1 1 6 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind full dna sequence full identification by DNA sequencing full dna sequence certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role bait bait physical association physical association The growth arrest and DNA damage-inducible protein, GADD34, was identified by its interaction with human inhibitor 1 (I-1), a protein kinase A (PKA)-activated inhibitor of type 1 protein serine/threonine phosphatase (PP1), in a yeast two-hybrid screen of a human brain cDNA library. ppp1r1a-ppp1r15a-2 2 5 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association The growth arrest and DNA damage-inducible protein, GADD34, was identified by its interaction with human inhibitor 1 (I-1), a protein kinase A (PKA)-activated inhibitor of type 1 protein serine/threonine phosphatase (PP1), in a yeast two-hybrid screen of a human brain cDNA library. Fig 1A ppp1r1a-gl7-1 2 4 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait physical association physical association we rescreened all I-1 interactors for their binding to yeast PP1 (GLC7). Fig 1A ppp1r1a-ppp1r15a-3 3 5 unspecified role unspecified role bait bait 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 c-terminal region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind full dna sequence full identification by DNA sequencing full dna sequence certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 21 23 physical association physical association To establish whether PP1 was required to recruit I-1 to GADD34 or whether the two proteins bound independently to GADD34, we expressed hexahistidine-tagged GADD34(233-674) and analyzed its association in vitro with recombinant human I-1 coupled to CNBr-activated Sepharose Figs. 2 and 3 mycrocyst-ppp1r15a-1 3 7 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey 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 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association HEK293T cell extracts were incubated with microcystin-LR-Sepharose (labeled Mcyst pulldown) to isolate cellular PP1/GADD34 complexes, and GADD34 was detected by Western immunoblotting Fig 4c ppp1r1a-ppp1r15a-4 3 5 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey 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 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. physical association physical association Immunoblotting the I-1-bound proteins with an anti-FLAG antibody established that all GADD34 polypeptides, including the full-length protein, were concentrated by the I-1 beads (Fig. 4D) Fig. 4D