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/ Dcs2, a novel stress-induced modulator of m7GpppX pyrophosphatase activity that locates to P bodies. Dcs2, a novel stress-induced modulator of m7GpppX pyrophosphatase activity that locates to P bodies. Journal of molecular biology 2006 imex curation Malys N., McCarthy JE., Malys N., McCarthy JE. john.mccarthy@manchester.ac.uk 05-Sep-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Malys N., McCarthy JE., Malys N., McCarthy JE. john.mccarthy@manchester.ac.uk Accepted 2017-AUG-31 AT 15:26 BST AT 15:26 BST by MGT Journal of molecular biology 2006 imex curation Its density gradient centrifugation where we only accept if pre-purified, we've always taken gel filtration. I've deleted the cleavage expt some of your points could have gone either way, so I decided it was more trouble than it was worth. Dcs2, a novel stress-induced modulator of m7GpppX pyrophosphatase activity that locates to P bodies. Dcs2, a novel stress-induced modulator of m7GpppX pyrophosphatase activity that locates to P bodies. Journal of molecular biology 2006 imex curation Malys N., McCarthy JE., Malys N., McCarthy JE. john.mccarthy@manchester.ac.uk 05-Sep-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom weight silver stain molecular weight estimation by silver staining weight silver stain Malys N., McCarthy JE., Malys N., McCarthy JE. john.mccarthy@manchester.ac.uk Accepted 2017-AUG-31 AT 15:26 BST AT 15:26 BST by MGT Journal of molecular biology 2006 imex curation Its density gradient centrifugation where we only accept if pre-purified, we've always taken gel filtration. I've deleted the cleavage expt some of your points could have gone either way, so I decided it was more trouble than it was worth. Dcs2, a novel stress-induced modulator of m7GpppX pyrophosphatase activity that locates to P bodies. Dcs2, a novel stress-induced modulator of m7GpppX pyrophosphatase activity that locates to P bodies. Journal of molecular biology 2006 imex curation Malys N., McCarthy JE., Malys N., McCarthy JE. john.mccarthy@manchester.ac.uk 05-Sep-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom weight by comassie molecular weight estimation by coomasie staining weight by comassie Malys N., McCarthy JE., Malys N., McCarthy JE. john.mccarthy@manchester.ac.uk Accepted 2017-AUG-31 AT 15:26 BST AT 15:26 BST by MGT Journal of molecular biology 2006 imex curation Its density gradient centrifugation where we only accept if pre-purified, we've always taken gel filtration. I've deleted the cleavage expt some of your points could have gone either way, so I decided it was more trouble than it was worth. Dcs2, a novel stress-induced modulator of m7GpppX pyrophosphatase activity that locates to P bodies. Dcs2, a novel stress-induced modulator of m7GpppX pyrophosphatase activity that locates to P bodies. Journal of molecular biology 2006 imex curation Malys N., McCarthy JE., Malys N., McCarthy JE. john.mccarthy@manchester.ac.uk 05-Sep-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Malys N., McCarthy JE., Malys N., McCarthy JE. john.mccarthy@manchester.ac.uk Accepted 2017-AUG-31 AT 15:26 BST AT 15:26 BST by MGT Journal of molecular biology 2006 imex curation Its density gradient centrifugation where we only accept if pre-purified, we've always taken gel filtration. I've deleted the cleavage expt some of your points could have gone either way, so I decided it was more trouble than it was worth. dcps_yeast m7GpppX diphosphatase DCS-1 Hint-related 7meGMP-directed hydrolase 1 DCS1 Scavenger mRNA-decapping enzyme DcpS YLR270W Protein Dcs1p protein protein yeast Saccharomyces cerevisiae Baker's yeast MSQLPTDFASLIKRFQFVSVLDSNPQTKVMSLLGTIDNKDAIITAEKTHFLFDETVRRPSQDGRSTPVLYNCENEYSCINGIQELKEITSNDIYYWGLSVIKQDMESNPTAKLNLIWPATPIHIKKYEQQNFHLVRETPEMYKRIVQPYIEEMCNNGRLKWVNNILYEGAESERVVYKDFSEENKDDGFLILPDMKWDGMNLDSLYLVAIVYRTDIKTIRDLRYSDRQWLINLNNKIRSIVPGCYNYAVHPDELRILVHYQPSYYHFHIHIVNIKHPGLGNSIAAGKAILLEDIIEMLNYLGPEGYMNKTITYAIGENHDLWKRGLEEELTKQLERDGIPKIPKIVNGFK 06A57C2344A3580A 06A57C2344A3580A dcs2_yeast Inactive diphosphatase DCS2 Histidine triad protein DCS2 Protein Dcs2p O3625 YOR173W protein protein yeast Saccharomyces cerevisiae Baker's yeast MGSQDLASLIGRFKYVRVLDSNPHTKVISLLGSIDGKDAVLTAEKTHFIFDETVRRPSQSGRSTPIFFHREIDEYSFLNGITDLKELTSNDIYYWGLSVLKQHILHNPTAKVNLIWPASQFHIKGYDQQDLHVVRETPDMYRNIVVPFIQEMCTSERMKWVNNILYEGAEDDRVVYKEYSSRNKEDGFVILPDMKWDGINIDSLYLVAIVYRDDIKSLRDLNPNHRDWLIRLNKKIKTIIPQHYDYNVNPDELRVFIHYQPSYYHFHVHIVNIRHPGVGEERGSGMTILLEDVIEALGFLGPEGYMKKTLTYVIGENHDLWKKGFKEEVEKQLKHDGIATSPEKGSGFNTNLG B76D89C0816019A4 B76D89C0816019A4 7-methyl-gtp 7-methyl-GTP small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) dcs2-dcs1-3 4 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait 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. 1A First, pull-down experiments using a FLAG-tagged version of Dcs2 revealed the accumulation of Dcs1:Dcs2 heterodimer after stress-induced synthesis of Dcs2 as cells run short of glucose Cells were glucoase-starved 7_methyl_gtp-dcs1-1 3 5 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association 80% of the bound Dcs1 could be specifically eluted by m7GpppG Fig. 1C dcs2-dcs1-4 2 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 3A dcs1-dcs2-5 2 5 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey physical association physical association Fig. 3A dcs2-dcs1-5 1 5 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 1B Cells were glucoase-starved dcs1-6 1 5 unspecified role unspecified role neutral component neutral component physical association physical association Fig. 1B