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/ SIRT1 regulates circadian clock gene expression through PER2 deacetylation. SIRT1 regulates circadian clock gene expression through PER2 deacetylation. Cell (0092-8674) 2008 imex curation Asher G., Gatfield D., Stratmann M., Reinke H., Dibner C., Kreppel F., Mostoslavsky R., Alt FW., Schibler U. Apoptosis - Interactions involving proteins with a function related to apoptosis ueli.schibler@molbio.unige.ch 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 12 mouse-liver Mus musculus liver liver anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Asher G., Gatfield D., Stratmann M., Reinke H., Dibner C., Kreppel F., Mostoslavsky R., Alt FW., Schibler U. Apoptosis - Interactions involving proteins with a function related to apoptosis Zeitgeber time - a standard of time based on the period of a zeitgeber (a stimulus capable of resetting a pacemaker or synchronizing a self-sustaining oscillation). Under standard light-dark cycles, the time of lights on usually defines zeitgeber time zero (ZT 0) for diurnal organisms and the time of lights off defines zeitgeber time twelve (ZT 12) for nocturnal animals. ueli.schibler@molbio.unige.ch Accepted 2008-SEP-20 by DAVET. 16-Oct-2008: Contacted by JYOTI. Zeitgeber time Cell (0092-8674) 2008 davet Only protein-protein interactions 12 imex curation SIRT1 regulates circadian clock gene expression through PER2 deacetylation. SIRT1 regulates circadian clock gene expression through PER2 deacetylation. Cell (0092-8674) 2008 imex curation Asher G., Gatfield D., Stratmann M., Reinke H., Dibner C., Kreppel F., Mostoslavsky R., Alt FW., Schibler U. Apoptosis - Interactions involving proteins with a function related to apoptosis ueli.schibler@molbio.unige.ch 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 12 mouse-liver Mus musculus liver liver anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Asher G., Gatfield D., Stratmann M., Reinke H., Dibner C., Kreppel F., Mostoslavsky R., Alt FW., Schibler U. Apoptosis - Interactions involving proteins with a function related to apoptosis ueli.schibler@molbio.unige.ch Accepted 2008-SEP-20 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 12 imex curation SIRT1 regulates circadian clock gene expression through PER2 deacetylation. SIRT1 regulates circadian clock gene expression through PER2 deacetylation. Cell (0092-8674) 2008 imex curation Asher G., Gatfield D., Stratmann M., Reinke H., Dibner C., Kreppel F., Mostoslavsky R., Alt FW., Schibler U. Apoptosis - Interactions involving proteins with a function related to apoptosis ueli.schibler@molbio.unige.ch 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 12 in vitro In vitro enzymatic study enzymatic study western blot western blot Immuno blot Immuno blot Asher G., Gatfield D., Stratmann M., Reinke H., Dibner C., Kreppel F., Mostoslavsky R., Alt FW., Schibler U. Apoptosis - Interactions involving proteins with a function related to apoptosis Additional information on SIRT1 taken from manufacturer's website. ueli.schibler@molbio.unige.ch Accepted 2008-SEP-20 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 12 imex curation SIRT1 regulates circadian clock gene expression through PER2 deacetylation. SIRT1 regulates circadian clock gene expression through PER2 deacetylation. Cell (0092-8674) 2008 imex curation Asher G., Gatfield D., Stratmann M., Reinke H., Dibner C., Kreppel F., Mostoslavsky R., Alt FW., Schibler U. Apoptosis - Interactions involving proteins with a function related to apoptosis ueli.schibler@molbio.unige.ch 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 12 mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Asher G., Gatfield D., Stratmann M., Reinke H., Dibner C., Kreppel F., Mostoslavsky R., Alt FW., Schibler U. Apoptosis - Interactions involving proteins with a function related to apoptosis ueli.schibler@molbio.unige.ch Accepted 2008-SEP-20 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 12 imex curation SIRT1 regulates circadian clock gene expression through PER2 deacetylation. SIRT1 regulates circadian clock gene expression through PER2 deacetylation. Cell (0092-8674) 2008 imex curation Asher G., Gatfield D., Stratmann M., Reinke H., Dibner C., Kreppel F., Mostoslavsky R., Alt FW., Schibler U. Apoptosis - Interactions involving proteins with a function related to apoptosis ueli.schibler@molbio.unige.ch 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 12 mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells confocal microscopy confocal microscopy immunostaining immunostaining Asher G., Gatfield D., Stratmann M., Reinke H., Dibner C., Kreppel F., Mostoslavsky R., Alt FW., Schibler U. Apoptosis - Interactions involving proteins with a function related to apoptosis ueli.schibler@molbio.unige.ch Accepted 2008-SEP-20 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 12 imex curation clock_mouse Circadian locomoter output cycles protein kaput Clock protein protein mouse Mus musculus Mouse MVFTVSCSKMSSIVDRDDSSIFDGLVEEDDKDKAKRVSRNKSEKKRRDQFNVLIKELGSMLPGNARKMDKSTVLQKSIDFLRKHKETTAQSDASEIRQDWKPTFLSNEEFTQLMLEALDGFFLAIMTDGSIIYVSESVTSLLEHLPSDLVDQSIFNFIPEGEHSEVYKILSTHLLESDSLTPEYLKSKNQLEFCCHMLRGTIDPKEPSTYEYVRFIGNFKSLTSVSTSTHNGFEGTIQRTHRPSYEDRVCFVATVRLATPQFIKEMCTVEEPNEEFTSRHSLEWKFLFLDHRAPPIIGYLPFEVLGTSGYDYYHVDDLENLAKCHEHLMQYGKGKSCYYRFLTKGQQWIWLQTHYYITYHQWNSRPEFIVCTHTVVSYAEVRAERRRELGIEESLPETAADKSQDSGSDNRINTVSLKEALERFDHSPTPSASSRSSRKSSHTAVSDPSSTPTKIPTDTSTPPRQHLPAHEKMTQRRSSFSSQSINSQSVGPSLTQPAMSQAANLPIPQGMSQFQFSAQLGAMQHLKDQLEQRTRMIEANIHRQQEELRKIQEQLQMVHGQGLQMFLQQSNPGLNFGSVQLSSGNSNIQQLTPVNMQGQVVPANQVQSGHISTGQHMIQQQTLQSTSTQQSQQSVMSGHSQQTSLPSQTPSTLTAPLYNTMVISQPAAGSMVQIPSSMPQNSTQSATVTTFTQDRQIRFSQGQQLVTKLVTAPVACGAVMVPSTMLMGQVVTAYPTFATQQQQAQTLSVTQQQQQQQQQPPQQQQQQQQSSQEQQLPSVQQPAQAQLGQPPQQFLQTSRLLHGNPSTQLILSAAFPLQQSTFPPSHHQQHQPQQQQQLPRHRTDSLTDPSKVQPQ DIP protein O08785 original sequence version: 149 DIP protein O08785-1 original sequence version: 138 DIP protein O08785 original sequence version: 138 DIP protein O08785 original sequence version: 108 DIP protein O08785 original sequence version: 143 DIP protein O08785 original sequence version: 155 DIP protein O08785 original sequence version: 159 DIP protein O08785 original sequence version: 160 9864D947049742F4 bmal1_mouse Aryl hydrocarbon receptor nuclear translocator-like protein 1 Arntl Bmal1 Brain and muscle ARNT-like 1 Arnt3 protein protein mouse Mus musculus Mouse MADQRMDISSTISDFMSPGPTDLLSGSLGTSGVDCNRKRKGSATDYQLDDFAFEESMDTDKDDPHGRLEYAEHQGRIKNAREAHSQIEKRRRDKMNSFIDELASLVPTCNAMSRKLDKLTVLRMAVQHMKTLRGATNPYTEANYKPTFLSDDELKHLILRAADGFLFVVGCDRGKILFVSESVFKILNYSQNDLIGQSLFDYLHPKDIAKVKEQLSSSDTAPRERLIDAKTGLPVKTDITPGPSRLCSGARRSFFCRMKCNRPSVKVEDKDFASTCSKKKDRKSFCTIHSTGYLKSWPPTKMGLDEDNEPDNEGCNLSCLVAIGRLHSHMVPQPANGEIRVKSMEYVSRHAIDGKFVFVDQRATAILAYLPQELLGTSCYEYFHQDDIGHLAECHRQVLQTREKITTNCYKFKIKDGSFITLRSRWFSFMNPWTKEVEYIVSTNTVVLANVLEGGDPTFPQLTAPPHSMDSMLPSGEGGPKRTHPTVPGIPGGTRAGAGKIGRMIAEEIMEIHRIRGSSPSSCGSSPLNITSTPPPDASSPGGKKILNGGTPDIPSTGLLPGQAQETPGYPYSDSSSILGENPHIGIDMIDNDQGSSSPSNDEAAMAVIMSLLEADAGLGGPVDFSDLPWPL DIP protein Q9WTL8 original sequence version: 134 DIP protein Q9WTL8 original sequence version: 123 DIP protein Q9WTL8 original sequence version: 94 DIP protein Q9WTL8 original sequence version: 128 DIP protein Q9WTL8 original sequence version: 144 DIP protein Q9WTL8 original sequence version: 145 9669C3712A95C2DE cry1_mouse Cryptochrome-1 Cry1 protein protein mouse Mus musculus Mouse MGVNAVHWFRKGLRLHDNPALKECIQGADTIRCVYILDPWFAGSSNVGINRWRFLLQCLEDLDANLRKLNSRLFVIRGQPADVFPRLFKEWNITKLSIEYDSEPFGKERDAAIKKLATEAGVEVIVRISHTLYDLDKIIELNGGQPPLTYKRFQTLVSKMEPLEMPADTITSDVIGKCMTPLSDDHDEKYGVPSLEELGFDTDGLSSAVWPGGETEALTRLERHLERKAWVANFERPRMNANSLLASPTGLSPYLRFGCLSCRLFYFKLTDLYKKVKKNSSPPLSLYGQLLWREFFYTAATNNPRFDKMEGNPICVQIPWDKNPEALAKWAEGRTGFPWIDAIMTQLRQEGWIHHLARHAVACFLTRGDLWISWEEGMKVFEELLLDADWSINAGSWMWLSCSSFFQQFFHCYCPVGFGRRTDPNGDYIRRYLPVLRGFPAKYIYDPWNAPEGIQKVAKCLIGVNYPKPMVNHAEASRLNIERMKQIYQQLSRYRGLGLLASVPSNSNGNGGLMGYAPGENVPSCSSSGNGGLMGYAPGENVPSCSGGNCSQGSGILHYAHGDSQQTHSLKQGRSSAGTGLSSGKRPSQEEDAQSVGPKVQRQSSN DIP protein P97784 original sequence version: 119 DIP protein P97784 original sequence version: 108 DIP protein P97784 original sequence version: 86 DIP protein P97784 original sequence version: 110 DIP protein P97784 original sequence version: 113 DIP protein P97784 original sequence version: 125 DIP protein P97784 original sequence version: 130 DIP protein P97784 original sequence version: 95 2F2B8DD53F0A9AF9 sir1_mouse NAD-dependent protein deacetylase sirtuin-1 NAD-dependent protein deacylase sirtuin-1 Sirt1 Sir2l1 SIR2alpha SIR2-like protein 1 Regulatory protein SIR2 homolog 1 protein protein mouse Mus musculus Mouse MADEVALALQAAGSPSAAAAMEAASQPADEPLRKRPRRDGPGLGRSPGEPSAAVAPAAAGCEAASAAAPAALWREAAGAAASAEREAPATAVAGDGDNGSGLRREPRAADDFDDDEGEEEDEAAAAAAAAAIGYRDNLLLTDGLLTNGFHSCESDDDDRTSHASSSDWTPRPRIGPYTFVQQHLMIGTDPRTILKDLLPETIPPPELDDMTLWQIVINILSEPPKRKKRKDINTIEDAVKLLQECKKIIVLTGAGVSVSCGIPDFRSRDGIYARLAVDFPDLPDPQAMFDIEYFRKDPRPFFKFAKEIYPGQFQPSLCHKFIALSDKEGKLLRNYTQNIDTLEQVAGIQRILQCHGSFATASCLICKYKVDCEAVRGDIFNQVVPRCPRCPADEPLAIMKPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHEVPQILINREPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRPQKELVHLSELPPTPLHISEDSSSPERTVPQDSSVIATLVDQATNNNVNDLEVSESSCVEEKPQEVQTSRNVENINVENPDFKAVGSSTADKNERTSVAETVRKCWPNRLAKEQISKRLEGNQYLFVPPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPLEDESEIEEFYNGLEDDTERPECAGGSGFGADGGDQEVVNEAIATRQELTDVNYPSDKS DIP protein Q923E4 original sequence version: 83 DIP protein Q923E4 original sequence version: 135 7F15625E29433119 per2_mouse Period circadian protein homolog 2 Per2 Circadian clock protein PERIOD 2 protein protein mouse Mus musculus Mouse MNGYVDFSPSPTSPTKEPGAPQPTQAVLQEDVDMSSGSSGNENCSTGRDSQGSDCDDNGKELRMLVESSNTHPSPDDAFRLMMTEAEHNPSTSGCSSEQSAKADAHKELIRTLKELKVHLPADKKAKGKASTLATLKYALRSVKQVKANEEYYQLLMSSESQPCSVDVPSYSMEQVEGITSEYIVKNADMFAVAVSLVSGKILYISNQVASIFHCKKDAFSDAKFVEFLAPHDVSVFHSYTTPYKLPPWSVCSGLDSFTQECMEEKSFFCRVSVGKHHENEIRYQPFRMTPYLVKVQEQQGAESQLCCLLLAERVHSGYEAPRIPPEKRIFTTTHTPNCLFQAVDERAVPLLGYLPQDLIETPVLVQLHPSDRPLMLAIHKKILQAGGQPFDYSPIRFRTRNGEYITLDTSWSSFINPWSRKISFIIGRHKVRVGPLNEDVFAAPPCPEEKTPHPSVQELTEQIHRLLMQPVPHSGSSGYGSLGSNGSHEHLMSQTSSSDSNGQEESHRRRSGIFKTSGKIQTKSHVSHESGGQKEASVAEMQSSPPAQVKAVTTIERDSSGASLPKASFPEELAYKNQPPCSYQQISCLDSVIRYLESCSEAATLKRKCEFPANIPSRKATVSPGLHSGEAARPSKVTSHTEVSAHLSSLTLPGKAESVVSLTSQCSYSSTIVHVGDKKPQPELETVEDMASGPESLDGAAGGLSQEKGPLQKLGLTKEVLAAHTQKEEQGFLQRFREVSRLSALQAHCQNYLQERSRAQASDRGLRNTSGLESSWKKTGKNRKLKSKRVKTRDSSESTGSGGPVSHRPPLMGLNATAWSPSDTSQSSCPSAPFPTAVPAYPLPVFQAPGIVSTPGTVVAPPAATHTGFTMPVVPMGTQPEFAVQPLPFAAPLAPVMAFMLPSYPFPPATPNLPQAFLPSQPHFPAHPTLASEITPASQAEFPSRTSTLRQPCACPVTPPAGTVALGRASPPLFQSRGSSPLQLNLLQLEEAPEGSTGAAGTLGTTGTAASGLDCTSGTSRDRQPKAPPTCNEPSDTQNSDAISTSSDLLNLLLGEDLCSATGSALSRSGASATSDSLGSSSLGFGTSQSGAGSSDTSHTSKYFGSIDSSENNHKAKMIPDTEESEQFIKYVLQDPIWLLMANTDDSIMMTYQLPSRDLQAVLKEDQEKLKLLQRSQPRFTEGQRRELREVHPWVHTGGLPTAIDVTGCVYCESEEKGNICLPYEEDSPSPGLCDTSEAKEEEGEQLTGPRIEAQT DIP protein O54943 original sequence version: 120 DIP protein O54943 original sequence version: 83 DIP protein O54943 original sequence version: 87 DIP protein O54943 original sequence version: 104 DIP protein O54943 original sequence version: 111 DIP protein O54943 original sequence version: 114 DIP protein O54943 original sequence version: 98 554B8AFF036CF7FB sir1_human NAD-dependent protein deacetylase sirtuin-1 NAD-dependent protein deacylase sirtuin-1 SIRT1 SIR2L1 SIR2-like protein 1 Regulatory protein SIR2 homolog 1 protein protein human Homo sapiens Human MADEAALALQPGGSPSAAGADREAASSPAGEPLRKRPRRDGPGLERSPGEPGGAAPEREVPAAARGCPGAAAAALWREAEAEAAAAGGEQEAQATAAAGEGDNGPGLQGPSREPPLADNLYDEDDDDEGEEEEEAAAAAIGYRDNLLFGDEIITNGFHSCESDEEDRASHASSSDWTPRPRIGPYTFVQQHLMIGTDPRTILKDLLPETIPPPELDDMTLWQIVINILSEPPKRKKRKDINTIEDAVKLLQECKKIIVLTGAGVSVSCGIPDFRSRDGIYARLAVDFPDLPDPQAMFDIEYFRKDPRPFFKFAKEIYPGQFQPSLCHKFIALSDKEGKLLRNYTQNIDTLEQVAGIQRIIQCHGSFATASCLICKYKVDCEAVRGDIFNQVVPRCPRCPADEPLAIMKPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHEVPQILINREPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRTQKELAYLSELPPTPLHVSEDSSSPERTSPPDSSVIVTLLDQAAKSNDDLDVSESKGCMEEKPQEVQTSRNVESIAEQMENPDLKNVGSSTGEKNERTSVAGTVRKCWPNRVAKEQISRRLDGNQYLFLPPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPMEDESEIEEFYNGLEDEPDVPERAGGAGFGTDGDDQEAINEAISVKQEVTDMNYPSNKS DIP protein Q96EB6 original sequence version: 140 DIP protein Q96EB6 original sequence version: 71 DIP protein Q96EB6 original sequence version: 125 DIP protein Q96EB6 original sequence version: 115 DIP protein Q96EB6 original sequence version: 63 DIP protein Q96EB6-1 original sequence version: 102 DIP protein Q96EB6 original sequence version: 102 DIP protein Q96EB6 original sequence version: 98 DIP protein Q96EB6 original sequence version: 144 DIP protein Q96EB6 original sequence version: 120 DIP protein Q96EB6-1 original sequence version: 120 DIP protein Q96EB6 original sequence version: 113 DIP protein Q96EB6-1 original sequence version: 117 2D3BEA6D73DA229F sirt1-clock-1 2 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait association association Both endogenous CLOCK and BMAL1 coimmunoprecipitated with SIRT1 in extracts obtained from mouse liver nuclei (Figure 5A) Fig. 5A sirt1-clock-2 4 9 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey association association Both endogenous CLOCK and BMAL1 coimmunoprecipitated with SIRT1 in extracts obtained from mouse liver nuclei (Figure 5A) and from cultured NIH 3T3 fibroblasts (Figure 5B). Fig. 5B sirt1-clock-3 5 6 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component colocalization colocalization coimmunostaining experiments for SIRT1 and CLOCK in NIH 3T3 cells showed that at least a fraction of these proteins colocalized in the nucleus (Figure 5C). Fig. 5C arntl1-clock-1 1 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey association association Fig. 5E ZT0 arntl1-clock-2 1 6 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey association association Fig. 5E ZT4 arntl1-clock-3 1 6 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey physical association physical association Fig. 5E ZT8 arntl1-clock-4 1 8 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey association association Fig. 5E ZT12 arntl1-clock-5 1 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait association association Fig. 5E ZT16 arntl1-clock-6 1 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait association association Fig. 5E ZT20 sirt1-clock-4 2 9 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association SIRT1 bound to CLOCK in a circadian manner with maximal binding around ZT4 (Figure 5F). Fig. 5F SIRT1 binds to CLOCK in a circadian manner with maximal binding around ZT4. sirt1-clock-5 4 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey association association Fig. 5G Interaction increased from 30 hours following cell synchronisation by Dexamethasone sirt1-per2-1 3 11 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) 10 enzyme target enzyme target substrate substrate 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 region naclys N-acetylated L-lysine NAcLys undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined mouse-3t3 Mus musculus embryo cells 3t3 Mouse embryo fibroblast cells deacetylation deacetylation reaction deacetylation Fig. 6B Reaction required NAD+ PER2 was acetylated (Figure S6) and that the acetyl groups were removed in vitro by recombinant SIRT1 in an NAD+-dependent manner (Figure 6B). Deacetylation of Per2