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/ Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. J. Biol. Chem. (0021-9258) 2006 imex curation King DA., Hall BE., Iwamoto MA., Win KZ., Chang JF., Ellenberger T. BioCreative - Critical Assessment of Information Extraction systems in Biology Chromatin - Epigenetic interactions resulting in chromatin modulation Bait protein, GST-Sir4-CC was immobilized on glutathione-conjugated multiwell plates were screened for binding against Sir3 fragment library. tome@biochem.wustl.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions 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 Accepted 2006-AUG-24 by DAVET. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. J. Biol. Chem. (0021-9258) 2006 imex curation King DA., Hall BE., Iwamoto MA., Win KZ., Chang JF., Ellenberger T. BioCreative - Critical Assessment of Information Extraction systems in Biology Chromatin - Epigenetic interactions resulting in chromatin modulation Bait protein, GST-Sir4-CC was immobilized on glutathione-conjugated multiwell plates were screened for binding against Sir3 fragment library. tome@biochem.wustl.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions 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 Accepted 2006-AUG-24 by DAVET. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. J. Biol. Chem. (0021-9258) 2006 imex curation King DA., Hall BE., Iwamoto MA., Win KZ., Chang JF., Ellenberger T. BioCreative - Critical Assessment of Information Extraction systems in Biology Chromatin - Epigenetic interactions resulting in chromatin modulation Bait protein, GST-Sir4-CC was immobilized on glutathione-conjugated multiwell plates were screened for binding against Sir3 fragment library. tome@biochem.wustl.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions 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 Accepted 2006-AUG-24 by DAVET. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. J. Biol. Chem. (0021-9258) 2006 imex curation King DA., Hall BE., Iwamoto MA., Win KZ., Chang JF., Ellenberger T. BioCreative - Critical Assessment of Information Extraction systems in Biology Chromatin - Epigenetic interactions resulting in chromatin modulation Bait protein, GST-Sir4-CC was immobilized on glutathione-conjugated multiwell plates were screened for binding against Sir3 fragment library. tome@biochem.wustl.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions 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 Accepted 2006-AUG-24 by DAVET. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. J. Biol. Chem. (0021-9258) 2006 imex curation King DA., Hall BE., Iwamoto MA., Win KZ., Chang JF., Ellenberger T. BioCreative - Critical Assessment of Information Extraction systems in Biology Chromatin - Epigenetic interactions resulting in chromatin modulation Bait protein, GST-Sir4-CC was immobilized on glutathione-conjugated multiwell plates were screened for binding against Sir3 fragment library. tome@biochem.wustl.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions 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 Accepted 2006-AUG-24 by DAVET. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation 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 CA2503D7645397AC 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 9C698765964F094E sir4-sir3 2 6 unspecified role unspecified role prey prey cat fusion protein fusion protein 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 ecoli Escherichia coli (strain K12) 7 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 c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P11978:p.Met1307Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain M N Wed May 02 15:06:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'met1307asn' Tue Jun 30 17:50:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Met1307Asn' ecoli Escherichia coli (strain K12) physical association physical association 2 These protein fragments bind specifically to Sir4-CC and fail to bind underivatized plates or a Sir4-CC(M1307N) point mutant that does not interact with full-length Sir3 sir4-sir3-1 5 7 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 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 ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey trx fusion protein fusion protein 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 region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction 3 Sir3 residues 464�522 are essential, but apparently not sufficient, for the interaction with Sir4. We conclude that Sir3-(464�728) is the minimal domain of Sir3 that stably interacts with Sir4. sir3-sir3 3 6 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 [EBI-995842]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 979-979, sequence length 978 [EBI-995842]833-979 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 6 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 4A The efficient association of Sir3-(832�978) with both Sir3 fragments shows that the dimerization function of Sir3 is mediated by C-terminal residues 832�978 and Sir4-binding residues 464�728. sir3-sir3-1 3 6 unspecified role unspecified role prey prey strep ii tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 6 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 region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-995866]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 979-979, sequence length 978 [EBI-995866]833-979 physical association physical association 4A The efficient association of Sir3-(832�978) with both Sir3 fragments shows that the dimerization function of Sir3 is mediated by C-terminal residues 832�978 and Sir4-binding residues 464�728. sir3-sir4 4 6 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 region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role fluorescence donor fluorescence donor tmr label tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction 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 4 6 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 [EBI-995989]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 979-979, sequence length 978 [EBI-995989]465-979 ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role fluorescence donor fluorescence donor tmr label tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction 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-sir3-2 5 6 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 [EBI-996004]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 979-979, sequence length 978 [EBI-996004]465-979 strep ii fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 7 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 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction 6A Both subunits of the Sir3 dimer are required to stabilize the complex with the Sir4 coiled-coil. Sir3 464-978 has the Sir4 interaction as well as Sir3 dimerisation domain. sir4-sir4 1 7 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 7 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 direct interaction direct interaction 7 Thus, the Sir4-CC domain alone or in complex with Sir3-(464�978) assembles into higher order protein arrays in the absence of DNA, albeit at high protein concentrations. Higher oligomers found. sir4-sir3-3 1 7 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 ecoli Escherichia coli (strain K12) 6 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 [EBI-996040]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 979-979, sequence length 978 [EBI-996040]465-979 ecoli Escherichia coli (strain K12) direct interaction direct interaction 7 Thus, the Sir4-CC domain alone or in complex with Sir3-(464�978) assembles into higher order protein arrays in the absence of DNA, albeit at high protein concentrations. Higher oligomers found.