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/ http://www.ebi.ac.uk/ king-2006-2 Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot King DA., Hall BE., Iwamoto MA., Win KZ., Chang JF., Ellenberger T. BioCreative - Critical Assessment of Information Extraction systems in Biology Bait protein, GST-Sir4-CC was immobilized on glutathione-conjugated multiwell plates were screened for binding against Sir3 fragment library. tome@biochem.wustl.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Chromatin - Epigenetic interactions resulting in chromatin modulation king-2006-3 Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast pull down pull down western blot western blot Immuno blot Immuno blot King DA., Hall BE., Iwamoto MA., Win KZ., Chang JF., Ellenberger T. BioCreative - Critical Assessment of Information Extraction systems in Biology Bait protein, GST-Sir4-CC was immobilized on glutathione-conjugated multiwell plates were screened for binding against Sir3 fragment library. tome@biochem.wustl.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Chromatin - Epigenetic interactions resulting in chromatin modulation king-2006-4 Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. in vitro In vitro fluorescence spectr classical fluorescence spectroscopy fluorescence spectr predetermined predetermined participant predetermined King DA., Hall BE., Iwamoto MA., Win KZ., Chang JF., Ellenberger T. BioCreative - Critical Assessment of Information Extraction systems in Biology Bait protein, GST-Sir4-CC was immobilized on glutathione-conjugated multiwell plates were screened for binding against Sir3 fragment library. tome@biochem.wustl.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Chromatin - Epigenetic interactions resulting in chromatin modulation king-2006-5 Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. in vitro In vitro dls dynamic light scattering dls predetermined predetermined participant predetermined King DA., Hall BE., Iwamoto MA., Win KZ., Chang JF., Ellenberger T. BioCreative - Critical Assessment of Information Extraction systems in Biology Bait protein, GST-Sir4-CC was immobilized on glutathione-conjugated multiwell plates were screened for binding against Sir3 fragment library. tome@biochem.wustl.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Chromatin - Epigenetic interactions resulting in chromatin modulation king-2006-1 Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. in vitro In vitro pull down pull down elisa enzyme linked immunosorbent assay ELISA ELISA elisa King DA., Hall BE., Iwamoto MA., Win KZ., Chang JF., Ellenberger T. BioCreative - Critical Assessment of Information Extraction systems in Biology Bait protein, GST-Sir4-CC was immobilized on glutathione-conjugated multiwell plates were screened for binding against Sir3 fragment library. tome@biochem.wustl.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Chromatin - Epigenetic interactions resulting in chromatin modulation sir4_yeast Regulatory protein SIR4 STE9 SIR4 ASD1 UTH2 YDR227W YD9934.12 Silent information regulator 4 protein protein yeast Saccharomyces cerevisiae Baker's yeast MPNDNKTPNRSSTPKFTKKPVTPNDKIPEREEKSNEVKTPKIPLFTFAKSKNYSRPSTAIHTSPHQPSDVKPTSHKQLQQPKSSPLKKNNYNSFPHSNLEKISNSKLLSLLRSKTSAGRIESNNPSHDASRSLASFEQTAFSRHAQQQTSTFNSKPVRTIVPISTSQTNNSFLSGVKSLLSEEKIRDYSKEILGINLANEQPVLEKPLKKGSADIGASVISLTKDKSIRKDTVEEKKEEKLNIGKNFAHSDSLSVPKVSAGDSGISPEESKARSPGIAKPNAIQTEVYGINEESTNERLEINQEKPVKLDENSANSTVASALDTNGTSATTETLTSKKIVPSPKKVAIDQDKITLHDEKTLAPSKHQPITSEQKMKEDADLKRMEILKSPHLSKSPADRPQGRRNSRNFSTRDEETTKLAFLVEYEGQENNYNSTSRSTEKKNDMNTSAKNKNGENKKIGKRPPEIMSTEAHVNKVTEETTKQIQSVRIDGRKVLQKVQGESHIDSRNNTLNVTPSKRPQLGEIPNPMKKHKPNEGRTPNISNGTINIQKKLEPKEIVRDILHTKESSNEAKKTIQNPLNKSQNTALPSTHKVTQKKDIKIGTNDLFQVESAPKISSEIDRENVKSKDEPVSKAVESKSLLNLFSNVLKAPFIKSESKPFSSDALSKEKANFLETIASTEKPENKTDKVSLSQPVSASKHEYSDNFPVSLSQPSKKSFANHTEDEQIEKKKICRGRMNTIITHPGKMELVYVSDSDDSSSDNDSLTDLESLSSGESNEIKVTNDLDTSAEKDQIQAGKWFDPVLDWRKSDRELTKNILWRIADKTTYDKETITDLIEQGIPKHSYLSGNPLTSVTNDICSVENYETSSAFFYQQVHKKDRLQYLPLYAVSTFENTNNTEKNDVTNKNINIGKHSQEQNSSSAKPSQIPTVSSPLGFEETKLSTTPTKSNRRVSHSDTNSSKPKNTKENLSKSSWRQEWLANLKLISVSLVDEFPSELSDSDRQIINEKMQLLKDIFANNLKSAISNNFRESDIIILKGEIEDYPMSSEIKIYYNELQNKPDAKKARFWSFMKTQRFVSNMGFDIQKSCEPVSISTSVKPHVVEPEHMADAKIMPKDILQITKKPLMVKNVKPSSPPDVKSLVQLSTMETKTLPEKKQFDSIFNSNKAKIIPGNGKHASENISLSFSRPASYGYFSVGKRVPIVEDRRVKQLDDITDSNTTEILTSVDVLGTHSQTGTQQSNMYTSTQKTELEIDNKDSVTECSKDMKEDGLSFVDIVLSKAASALDEKEKQLAVANEIIRSLSDEVMRNEIRITSLQGDLTFTKKCLENARSQISEKDAKINKLMEKDFQVNKEIKPY 9C698765964F094E sir3_yeast Regulatory protein SIR3 SIR3 CMT1 MAR2 STE8 L9753.10 YLR442C Silent information regulator 3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MAKTLKDLDGWQVIITDDQGRVIDDNNRRRSRKRGGENVFLKRISDGLSFGKGESVIFNDNVTETYSVYLIHEIRLNTLNNVVEIWVFSYLRWFELKPKLYYEQFRPDLIKEDHPLEFYKDKFFNEVNKSELYLTAELSEIWLKDFIAVGQILPESQWNDSSIDKIEDRDFLVRYACEPTAEKFVPIDIFQIIRRVKEMEPKQSDEYLKRVSVPVSGQKTNRQVMHKMGVERSSKRLAKKPSMKKIKIEPSADDDVNNGNIPSQRGTSTTHGSISPQEESVSPNISSASPSALTSPTDSSKILQKRSISKELIVSEEIPINSSEQESDYEPNNETSVLSSKPGSKPEKTSTELVDGRENFVYANNPEVSDDGGLEEETDEVSSESSDEAIIPVNKRRGAHGSELSSKIRKIHIQETQEFSKNYTTETDNEMNGNGKPGIPRGNTKIHSMNENPTPEKGNAKMIDFATLSKLKKKYQIILDRFAPDNQVTDSSQLNKLTDEQSSLDVAGLEDKFRKACSSSGRETILSNFNADINLEESIRESLQKRELLKSQVEDFTRIFLPIYDSLMSSQNKLFYITNADDSTKFQLVNDVMDELITSSARKELPIFDYIHIDALELAGMDALYEKIWFAISKENLCGDISLEALNFYITNVPKAKKRKTLILIQNPENLLSEKILQYFEKWISSKNSKLSIICVGGHNVTIREQINIMPSLKAHFTEIKLNKVDKNELQQMIITRLKSLLKPFHVKVNDKKEMTIYNNIREGQNQKIPDNVIVINHKINNKITQLIAKNVANVSGSTEKAFKICEAAVEISKKDFVRKGGLQKGKLVVSQEMVPRYFSEAINGFKDETISKKIIGMSLLMRTFLYTLAQETEGTNRHTLALETVLIKMVKMLRDNPGYKASKEIKKVICGAWEPAITIEKLKQFSWISVVNDLVGEKLVVVVLEEPSASIMVELKLPLEINYAFSMDEEFKNMDCI CA2503D7645397AC sir4-sir3-1 498894 N/A 498899 unspecified role unspecified role bait bait c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) N/A 498900 unspecified role unspecified role prey prey trx fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 3 sir4-sir3-2 498894 N/A 498900 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false strep ii fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 498899 unspecified role unspecified role bait bait c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 6A Sir3 464-978 has the Sir4 interaction as well as Sir3 dimerisation domain. sir3-sir3 498895 N/A 498900 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false 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 false N/A 498900 unspecified role unspecified role prey prey 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 false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false 4A sir3-sir3-1 498895 N/A 498900 unspecified role unspecified role prey prey strep ii tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 498900 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 false region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false 4A sir3-sir4 498896 N/A 498900 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 498899 unspecified role unspecified role fluorescence donor fluorescence donor tmr label tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 5 and table1 The cooperative binding of Sir3-Sir4 was observed. An antiparallel association of the Sir3 dimers was also observed suggestive of a self-assembly process capable of generating higher order protein complexes sir3-sir4-1 498896 N/A 498900 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 498899 unspecified role unspecified role fluorescence donor fluorescence donor tmr label tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 5, 6B and table 1 The cooperative binding of Sir3-Sir4 was observed. An antiparallel association of the Sir3 dimers was also observed suggestive of a self-assembly process capable of generating higher order protein complexes In this assay the dimers of Sir3 interact with Sir4 construct and the concentration of Sir4 required to rech saturation was high. Sir4 binding activity being limited to the Sir3-(464�978) dimers constituting one-fourth of the total dimers in this mixture of subunits. sir4-sir4 498897 N/A 498899 unspecified role unspecified role neutral component neutral component c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 498899 unspecified role unspecified role neutral component neutral component c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false 7 Higher oligomers found. sir4-sir3-3 498897 N/A 498899 unspecified role unspecified role neutral component neutral component c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 498900 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 7 Higher oligomers found. sir4-sir3 498898 N/A 498900 unspecified role unspecified role prey prey cat fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 498899 unspecified role unspecified role bait bait 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 false c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P11978:p.Met1307Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false 2