MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it SIRT1 is regulated by a PPAR{γ}-SIRT1 negative feedback loop associated with senescence. SIRT1 is regulated by a PPAR{γ}-SIRT1 negative feedback loop associated with senescence. Nucleic acids research 2010 imex curation Han L., Zhou R., Niu J., McNutt MA., Wang P., Tong T. supplementary materials are not retrievable, so cannot be added nor checked. ttj@bjmu.edu.cn 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Han L., Zhou R., Niu J., McNutt MA., Wang P., Tong T. supplementary materials are not retrievable, so cannot be added nor checked. ttj@bjmu.edu.cn Accepted 2018-OCT-15 AT 17:19 BST AT 17:19 BST by LIVIAP Nucleic acids research 2010 imex curation fatto! SIRT1 is regulated by a PPAR{γ}-SIRT1 negative feedback loop associated with senescence. SIRT1 is regulated by a PPAR{γ}-SIRT1 negative feedback loop associated with senescence. Nucleic acids research 2010 imex curation Han L., Zhou R., Niu J., McNutt MA., Wang P., Tong T. supplementary materials are not retrievable, so cannot be added nor checked. ttj@bjmu.edu.cn 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Han L., Zhou R., Niu J., McNutt MA., Wang P., Tong T. supplementary materials are not retrievable, so cannot be added nor checked. ttj@bjmu.edu.cn Accepted 2018-OCT-15 AT 17:19 BST AT 17:19 BST by LIVIAP Nucleic acids research 2010 imex curation fatto! SIRT1 is regulated by a PPAR{γ}-SIRT1 negative feedback loop associated with senescence. SIRT1 is regulated by a PPAR{γ}-SIRT1 negative feedback loop associated with senescence. Nucleic acids research 2010 imex curation Han L., Zhou R., Niu J., McNutt MA., Wang P., Tong T. supplementary materials are not retrievable, so cannot be added nor checked. ttj@bjmu.edu.cn 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Han L., Zhou R., Niu J., McNutt MA., Wang P., Tong T. supplementary materials are not retrievable, so cannot be added nor checked. Nuclear extract were used in this experiment ttj@bjmu.edu.cn Accepted 2018-OCT-15 AT 17:19 BST AT 17:19 BST by LIVIAP Nucleic acids research 2010 imex curation fatto! 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 pparg_human Peroxisome proliferator-activated receptor gamma Nuclear receptor subfamily 1 group C member 3 PPARG NR1C3 protein protein human Homo sapiens Human MGETLGDSPIDPESDSFTDTLSANISQEMTMVDTEMPFWPTNFGISSVDLSVMEDHSHSFDIKPFTTVDFSSISTPHYEDIPFTRTDPVVADYKYDLKLQEYQSAIKVEPASPPYYSEKTQLYNKPHEEPSNSLMAIECRVCGDKASGFHYGVHACEGCKGFFRRTIRLKLIYDRCDLNCRIHKKSRNKCQYCRFQKCLAVGMSHNAIRFGRMPQAEKEKLLAEISSDIDQLNPESADLRALAKHLYDSYIKSFPLTKAKARAILTGKTTDKSPFVIYDMNSLMMGEDKIKFKHITPLQEQSKEVAIRIFQGCQFRSVEAVQEITEYAKSIPGFVNLDLNDQVTLLKYGVHEIIYTMLASLMNKDGVLISEGQGFMTREFLKSLRKPFGDFMEPKFEFAVKFNALELDDSDLAIFIAVIILSGDRPGLLNVKPIEDIQDNLLQALELQLKLNHPESSQLFAKLLQKMTDLRQIVTEHVQLLQVIKKTETDMSLHPLLQEIYKDLY 3933EFF36A0E4CAF ep300_human Histone acetyltransferase p300 Histone butyryltransferase p300 Histone crotonyltransferase p300 Protein propionyltransferase p300 EP300 P300 E1A-associated protein p300 Protein 2-hydroxyisobutyryltransferase p300 Protein lactyltransferas p300 protein protein human Homo sapiens Human MAENVVEPGPPSAKRPKLSSPALSASASDGTDFGSLFDLEHDLPDELINSTELGLTNGGDINQLQTSLGMVQDAASKHKQLSELLRSGSSPNLNMGVGGPGQVMASQAQQSSPGLGLINSMVKSPMTQAGLTSPNMGMGTSGPNQGPTQSTGMMNSPVNQPAMGMNTGMNAGMNPGMLAAGNGQGIMPNQVMNGSIGAGRGRQNMQYPNPGMGSAGNLLTEPLQQGSPQMGGQTGLRGPQPLKMGMMNNPNPYGSPYTQNPGQQIGASGLGLQIQTKTVLSNNLSPFAMDKKAVPGGGMPNMGQQPAPQVQQPGLVTPVAQGMGSGAHTADPEKRKLIQQQLVLLLHAHKCQRREQANGEVRQCNLPHCRTMKNVLNHMTHCQSGKSCQVAHCASSRQIISHWKNCTRHDCPVCLPLKNAGDKRNQQPILTGAPVGLGNPSSLGVGQQSAPNLSTVSQIDPSSIERAYAALGLPYQVNQMPTQPQVQAKNQQNQQPGQSPQGMRPMSNMSASPMGVNGGVGVQTPSLLSDSMLHSAINSQNPMMSENASVPSLGPMPTAAQPSTTGIRKQWHEDITQDLRNHLVHKLVQAIFPTPDPAALKDRRMENLVAYARKVEGDMYESANNRAEYYHLLAEKIYKIQKELEEKRRTRLQKQNMLPNAAGMVPVSMNPGPNMGQPQPGMTSNGPLPDPSMIRGSVPNQMMPRITPQSGLNQFGQMSMAQPPIVPRQTPPLQHHGQLAQPGALNPPMGYGPRMQQPSNQGQFLPQTQFPSQGMNVTNIPLAPSSGQAPVSQAQMSSSSCPVNSPIMPPGSQGSHIHCPQLPQPALHQNSPSPVPSRTPTPHHTPPSIGAQQPPATTIPAPVPTPPAMPPGPQSQALHPPPRQTPTPPTTQLPQQVQPSLPAAPSADQPQQQPRSQQSTAASVPTPTAPLLPPQPATPLSQPAVSIEGQVSNPPSTSSTEVNSQAIAEKQPSQEVKMEAKMEVDQPEPADTQPEDISESKVEDCKMESTETEERSTELKTEIKEEEDQPSTSATQSSPAPGQSKKKIFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDIVKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQEIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQGESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAGFVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHASDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPPPNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDYIFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFEGDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLSRGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDLMDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQDRFVYTCNECKHHVETRWHCTVCEDYDLCITCYNTKNHDHKMEKLGLGLDDESNNQQAAATQSPGDSRRLSIQRCIQSLVHACQCRNANCSLPSCQKMKRVVQHTKGCKRKTNGGCPICKQLIALCCYHAKHCQENKCPVPFCLNIKQKLRQQQLQHRLQQAQMLRRRMASMQRTGVVGQQQGLPSPTPATPTTPTGQQPTTPQTPQPTSQPQPTPPNSMPPYLPRTQAAGPVSQGKAAGQVTPPTPPQTAQPPLPGPPPAAVEMAMQIQRAAETQRQMAHVQIFQRPIQHQMPPMTPMAPMGMNPPPMTRGPSGHLEPGMGPTGMQQQPPWSQGGLPQPQQLQSGMPRPAMMSVAQHGQPLNMAPQPGLGQVGISPLKPGTVSQQALQNLLRTLRSPSSPLQQQQVLSILHANPQLLAAFIKQRAAKYANSNPQPIPGQPGMPQGQPGLQPPTMPGQQGVHSNPAMQNMNPMQAGVQRAGLPQQQPQQQLQPPMGGMSPQAQQMNMNHNTMPSQFRDILRRQQMMQQQQQQGAGPGIGPGMANHNQFQQPQGVGYPPQQQQRMQHHMQQMQQGNMGQIGQLPQALGAEAGASLQAYQQRLLQQQMGSPVQPNPMSPQQHMLPNQAQSPHLQGQQIPNSLSNQVRSPQPVPSPRPQSQPPHSSPSPRMQPQPSPHHVSPQTSSPHPGLVAAQANPMEQGHFASPDQNSMLSQLASNPGMANLHGASATDLGLSTDNSDLNSNLSQSTLDIH DIP protein Q09472 original sequence version: 193 DIP protein Q09472 original sequence version: 186 DIP protein Q09472 original sequence version: 107 DIP protein Q09472 original sequence version: 112 DIP protein Q09472 original sequence version: 181 DIP protein Q09472 original sequence version: 183 DIP protein Q09472 original sequence version: 136 DIP protein Q09472 original sequence version: 146 DIP protein Q09472 original sequence version: 165 DIP protein Q09472 original sequence version: 192 DIP protein Q09472 original sequence version: 200 DIP protein Q09472 original sequence version: 206 DIP protein Q09472 original sequence version: 194 DIP protein Q09472 original sequence version: 171 DIP protein Q09472 original sequence version: 142 DIP protein Q09472 original sequence version: 180 8E869E1F174A6FEB sir1_human_gene SIRT1 Human Gene gene gene human Homo sapiens Human pparg-sir1_human_gene-1 3 5 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f4a Using the ChIP assay, we found that endogenously expressed PPARγ specifically binds to the PPRE cluster on the SIRT1 promoter in HeLa cells ( Figure 4 A). Using the ChIP assay, we found that endogenously expressed PPARγ specifically binds to the PPRE cluster on the SIRT1 promoter in HeLa cells ( Figure 4 A). sir1_human_gene-sirt1-1 3 4 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f4a Indeed, the ChIP assay using SIRT1 antibody indicated that endogenous SIRT1 was present on its own promoter in HeLa cells ( Figure 4 A). Indeed, the ChIP assay using SIRT1 antibody indicated that endogenous SIRT1 was present on its own promoter in HeLa cells ( Figure 4 A). pparg-10 2 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tag flag tag 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 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tag myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region binding region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Our results showed that the SIRT1 catalytic domain is required for PPARγ to recruit SIRT1 ( Figure 3 B). Collectively, these data indicate that the SIRT1 catalytic domain is necessary and sufficient for the interaction of PPARγ and SIRT1. Our results showed that the SIRT1 catalytic domain is required for PPARγ to recruit SIRT1 ( Figure 3 B). Collectively, these data indicate that the SIRT1 catalytic domain is necessary and sufficient for the interaction of PPARγ and SIRT1. f3b pparg-sirt1-1 1 5 unspecified role unspecified role bait bait purified purified gst tag gst tag 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 4 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated myc tag myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association To confirm that this reflected a direct physical interaction, we employed an in vitro interaction assay with recombinant GST–PPARγ. Based on the result of this GST pull-down assay ( Figure 3 A), SIRT1 interacts directly with PPARγ in vitro , which raises the possibility that PPARγ may be a substrate for SIRT1 deacetylase. To confirm that this reflected a direct physical interaction, we employed an in vitro interaction assay with recombinant GST–PPARγ. Based on the result of this GST pull-down assay ( Figure 3 A), SIRT1 interacts directly with PPARγ in vitro , which raises the possibility that PPARγ may be a substrate for SIRT1 deacetylase. f3a pparg-sirt1-2 2 5 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell flag tag flag tag 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 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell myc tag myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region binding region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Our results showed that the SIRT1 catalytic domain is required for PPARγ to recruit SIRT1 ( Figure 3 B). Collectively, these data indicate that the SIRT1 catalytic domain is necessary and sufficient for the interaction of PPARγ and SIRT1. Our results showed that the SIRT1 catalytic domain is required for PPARγ to recruit SIRT1 ( Figure 3 B). Collectively, these data indicate that the SIRT1 catalytic domain is necessary and sufficient for the interaction of PPARγ and SIRT1. f3b sirt1-pparg-1 3 5 unspecified role unspecified role prey prey partially purified partially purified 4 unspecified role unspecified role bait bait partially purified partially purified 7 unspecified role unspecified role prey prey partially purified partially purified association association f4d In both of these ChIP-upon-ChIP assays, endogenous PPARg and SIRT1 co-localized on the 50 SIRT1 promoter site, suggesting that PPARg recruits SIRT1 to SIRT1 promoter (Figure 4D). sirt1-pparg-3 3 6 unspecified role unspecified role prey prey partially purified partially purified 4 unspecified role unspecified role bait bait partially purified partially purified 7 unspecified role unspecified role prey prey association association f5b It is also recognized that protein acetylation is generally mediated by acetyltransferases such as the protein p300, and SIRT1 and p300 can interact (24) (also Supplementary Data, Figure 4) to form a complex and bind the SIRT1 promoter (Figure 5B). sirt1-pparg-4 3 5 unspecified role unspecified role bait bait partially purified partially purified 4 unspecified role unspecified role prey prey partially purified partially purified 7 unspecified role unspecified role prey prey partially purified partially purified association association f4d In both of these ChIP-upon-ChIP assays, endogenous PPARg and SIRT1 co-localized on the 50 SIRT1 promoter site, suggesting that PPARg recruits SIRT1 to SIRT1 promoter (Figure 4D). ep300-sir1_human_gene-1 3 7 unspecified role unspecified role prey prey partially purified partially purified 6 unspecified role unspecified role bait bait partially purified partially purified 4 unspecified role unspecified role prey prey partially purified partially purified physical association physical association f5b It is also recognized that protein acetylation is generally mediated by acetyltransferases such as the protein p300, and SIRT1 and p300 can interact (24) (also Supplementary Data, Figure 4) to form a complex and bind the SIRT1 promoter (Figure 5B).