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/ SIRT2 regulates NF-κB dependent gene expression through deacetylation of p65 Lys310. SIRT2 regulates NF-κB dependent gene expression through deacetylation of p65 Lys310. J. Cell. Sci. (0021-9533) 2010 imex curation Rothgiesser K.M., Erener S., Waibel S., Lüscher B., Hottiger M.O. IBD - Inflammatory bowel disease The authors also show acetylation of p65 at lys-310 by p300 in an in vitro acetylation assay; however, the origin of the p300 construct used in this experiment could not be established, hence this interaction is not captured here. Mammalian expression vectors for human SIRT cDNA's are described in PMID:16079181, while those for human p65 and p65 mutants are described in PMID:18263619. HA-tagged SIRT2 is described in PMID:18332217. hottiger@vetbio.uzh.ch 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-thp-1 Homo sapiens acute monocytic leukemia cells thp-1 Human acute monocytic leukemia cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Rothgiesser K.M., Erener S., Waibel S., Lüscher B., Hottiger M.O. IBD - Inflammatory bowel disease The authors also show acetylation of p65 at lys-310 by p300 in an in vitro acetylation assay; however, the origin of the p300 construct used in this experiment could not be established, hence this interaction is not captured here. Mammalian expression vectors for human SIRT cDNA's are described in PMID:16079181, while those for human p65 and p65 mutants are described in PMID:18263619. HA-tagged SIRT2 is described in PMID:18332217. hottiger@vetbio.uzh.ch Accepted 2014-DEC-17 by ORCHARD J. Cell. Sci. (0021-9533) 2010 Only protein-protein interactions imex curation SIRT2 regulates NF-κB dependent gene expression through deacetylation of p65 Lys310. SIRT2 regulates NF-κB dependent gene expression through deacetylation of p65 Lys310. J. Cell. Sci. (0021-9533) 2010 imex curation Rothgiesser K.M., Erener S., Waibel S., Lüscher B., Hottiger M.O. IBD - Inflammatory bowel disease The authors also show acetylation of p65 at lys-310 by p300 in an in vitro acetylation assay; however, the origin of the p300 construct used in this experiment could not be established, hence this interaction is not captured here. Mammalian expression vectors for human SIRT cDNA's are described in PMID:16079181, while those for human p65 and p65 mutants are described in PMID:18263619. HA-tagged SIRT2 is described in PMID:18332217. hottiger@vetbio.uzh.ch 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro deacetylase assay deacetylase assay western blot western blot Immuno blot Immuno blot Rothgiesser K.M., Erener S., Waibel S., Lüscher B., Hottiger M.O. IBD - Inflammatory bowel disease The authors also show acetylation of p65 at lys-310 by p300 in an in vitro acetylation assay; however, the origin of the p300 construct used in this experiment could not be established, hence this interaction is not captured here. Prior to the in vitro deacetylation assay, recombinant p65 was acetylated in vitro by incubation with recombinant human p300 (as described in PMID:11430825). Mammalian expression vectors for human SIRT cDNA's are described in PMID:16079181, while those for human p65 and p65 mutants are described in PMID:18263619. HA-tagged SIRT2 is described in PMID:18332217. hottiger@vetbio.uzh.ch Accepted 2014-DEC-17 by ORCHARD J. Cell. Sci. (0021-9533) 2010 Only protein-protein interactions imex curation SIRT2 regulates NF-κB dependent gene expression through deacetylation of p65 Lys310. SIRT2 regulates NF-κB dependent gene expression through deacetylation of p65 Lys310. J. Cell. Sci. (0021-9533) 2010 imex curation Rothgiesser K.M., Erener S., Waibel S., Lüscher B., Hottiger M.O. IBD - Inflammatory bowel disease The authors also show acetylation of p65 at lys-310 by p300 in an in vitro acetylation assay; however, the origin of the p300 construct used in this experiment could not be established, hence this interaction is not captured here. Mammalian expression vectors for human SIRT cDNA's are described in PMID:16079181, while those for human p65 and p65 mutants are described in PMID:18263619. HA-tagged SIRT2 is described in PMID:18332217. hottiger@vetbio.uzh.ch 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 confocal microscopy confocal microscopy immunocytochemistry immunocytochemistry Rothgiesser K.M., Erener S., Waibel S., Lüscher B., Hottiger M.O. IBD - Inflammatory bowel disease The authors also show acetylation of p65 at lys-310 by p300 in an in vitro acetylation assay; however, the origin of the p300 construct used in this experiment could not be established, hence this interaction is not captured here. Mammalian expression vectors for human SIRT cDNA's are described in PMID:16079181, while those for human p65 and p65 mutants are described in PMID:18263619. HA-tagged SIRT2 is described in PMID:18332217. hottiger@vetbio.uzh.ch Accepted 2014-DEC-17 by ORCHARD J. Cell. Sci. (0021-9533) 2010 Only protein-protein interactions imex curation sir2_human NAD-dependent protein deacetylase sirtuin-2 SIR2-like protein 2 SIRT2 SIR2L SIR2L2 Regulatory protein SIR2 homolog 2 protein protein human Homo sapiens Human MAEPDPSHPLETQAGKVQEAQDSDSDSEGGAAGGEADMDFLRNLFSQTLSLGSQKERLLDELTLEGVARYMQSERCRRVICLVGAGISTSAGIPDFRSPSTGLYDNLEKYHLPYPEAIFEISYFKKHPEPFFALAKELYPGQFKPTICHYFMRLLKDKGLLLRCYTQNIDTLERIAGLEQEDLVEAHGTFYTSHCVSASCRHEYPLSWMKEKIFSEVTPKCEDCQSLVKPDIVFFGESLPARFFSCMQSDFLKVDLLLVMGTSLQVQPFASLISKAPLSTPRLLINKEKAGQSDPFLGMIMGLGGGMDFDSKKAYRDVAWLGECDQGCLALAELLGWKKELEDLVRREHASIDAQSGAGVPNPSTSASPKKSPPPAKDEARTTEREKPQ DIP protein Q8IXJ6 original sequence version: 113 A392442A8F6316F1 tf65_human Transcription factor p65 Nuclear factor NF-kappa-B p65 subunit RELA NFKB3 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 protein protein human Homo sapiens Human MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGSIPGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAELCPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQVTVRDPSGRPLRLPPVLSHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQKEDIEVYFTGPGWEARGSFSQADVHRQVAIVFRTPPYADPSLQAPVRVSMQLRRPSDRELSEPMEFQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISS DIP protein Q04206 original sequence version: 170 DIP protein Q04206 original sequence version: 173 DIP protein Q04206 original sequence version: 99 DIP protein Q04206-1 original sequence version: 99 DIP protein Q04206 original sequence version: 168 DIP protein Q04206 original sequence version: 128 DIP protein Q04206 original sequence version: 131 DIP protein Q04206 original sequence version: 153 DIP protein Q04206 original sequence version: 185 DIP protein Q04206 original sequence version: 181 DIP protein Q04206 original sequence version: 183 8147A6AF9F2445C6 q8ixj6-2 NAD-dependent protein deacetylase sirtuin-2 SIRT2 SIR2L SIR2L2 SIR2-like protein 2 Regulatory protein SIR2 homolog 2 protein protein human Homo sapiens Human MDFLRNLFSQTLSLGSQKERLLDELTLEGVARYMQSERCRRVICLVGAGISTSAGIPDFRSPSTGLYDNLEKYHLPYPEAIFEISYFKKHPEPFFALAKELYPGQFKPTICHYFMRLLKDKGLLLRCYTQNIDTLERIAGLEQEDLVEAHGTFYTSHCVSASCRHEYPLSWMKEKIFSEVTPKCEDCQSLVKPDIVFFGESLPARFFSCMQSDFLKVDLLLVMGTSLQVQPFASLISKAPLSTPRLLINKEKAGQSDPFLGMIMGLGGGMDFDSKKAYRDVAWLGECDQGCLALAELLGWKKELEDLVRREHASIDAQSGAGVPNPSTSASPKKSPPPAKDEARTTEREKPQ FFED07DEF9E3416A h4_human Histone H4 H4-16 H4C1 H4C11 H4C12 H4C13 H4C14 H4C15 H4C2 H4C3 H4C4 H4C5 H4C6 H4C8 H4C9 H4/A H4/B H4/C H4/D H4/E H4/G H4/H H4/I H4/J H4/K H4/M H4/N H4/O H4F2 H4FA H4FB H4FC H4FD H4FE H4FG H4FH H4FI H4FJ H4FK H4FM H4FN H4FO HIST1H4A HIST1H4B HIST1H4C HIST1H4D HIST1H4E HIST1H4F HIST1H4H HIST1H4I HIST1H4J HIST1H4K HIST1H4L HIST2H4 HIST2H4A HIST2H4B HIST4H4 protein protein human Homo sapiens Human MSGRGKGGKGLGKGGAKRHRKVLRDNIQGITKPAIRRLARRGGVKRISGLIYEETRGVLKVFLENVIRDAVTYTEHAKRKTVTAMDVVYALKRQGRTLYGFGG DIP protein P62805 original sequence version: 68 DIP protein P62805 original sequence version: 111 DIP protein P62805 original sequence version: 94 DIP protein P62805 original sequence version: 97 DIP protein P62805 original sequence version: 116 DIP protein P62805 original sequence version: 105 DIP protein P62805 original sequence version: 120 DIP protein P62805 original sequence version: 144 DIP protein P62805 original sequence version: 150 DIP protein P62805 original sequence version: 154 DIP protein P62805 original sequence version: 156 DIP protein P62805 original sequence version: 112 DIP protein P62805 original sequence version: 121 DIP protein P62805 original sequence version: 123 DIP protein P62805 original sequence version: 126 DIP protein P62805 original sequence version: 132 DIP protein P62805 original sequence version: 136 DIP protein P62805 original sequence version: 119 A9E5DFD3F8B97598 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 rela-sirt2 2 4 enzyme enzyme neutral component neutral component c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range spofr-sf_21 Spodoptera frugiperda insect cells sf_21 Insect cells derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae. full name: IPLB-SF-21-AE cells are susceptible to Baculovirus infection. 5 enzyme target enzyme target substrate substrate neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined required acetyllys-310 naclys N-acetylated L-lysine NAcLys certain certain sequence position certain certain certain sequence position certain required acetyllys-314 naclys N-acetylated L-lysine NAcLys certain certain sequence position certain certain certain sequence position certain required acetyllys-315 naclys N-acetylated L-lysine NAcLys certain certain sequence position certain certain certain sequence position certain spofr-sf_21 Spodoptera frugiperda insect cells sf_21 Insect cells derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae. full name: IPLB-SF-21-AE cells are susceptible to Baculovirus infection. deacetylation deacetylation reaction deacetylation Fig. 2b, Suppl. Fig. S1a, S1e NAD+ (CHEBI:15846) SIRT2 deacetylated p65 in vitro at lysine-310, lysine-314 and lysine-315. Q8IXJ6 is required for deacetylation of Q04206 Q8IXJ6 decreases activity of Q04206 CHEBI:15846 increases activity of Q8IXJ6 P01375 increases binding of Q8IXJ6 to Q04206 rela-sirt1-4 2 8 enzyme enzyme neutral component neutral component c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range spofr-sf_21 Spodoptera frugiperda insect cells sf_21 Insect cells derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae. full name: IPLB-SF-21-AE cells are susceptible to Baculovirus infection. 5 enzyme target enzyme target substrate substrate neutral component neutral component required acetyllys-310 naclys N-acetylated L-lysine NAcLys certain certain sequence position certain certain certain sequence position certain required acetyllys-315 naclys N-acetylated L-lysine NAcLys certain certain sequence position certain certain certain sequence position certain region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined spofr-sf_21 Spodoptera frugiperda insect cells sf_21 Insect cells derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae. full name: IPLB-SF-21-AE cells are susceptible to Baculovirus infection. deacetylation deacetylation reaction deacetylation Suppl. Fig. S1a, S1e SIRT1 deacetylated p65 in vitro at lysine-310 and lysine-315. While p65 was deacetylated in vitro at lysine-310 by SIRT2 after 30 minutes, deacetylation by SIRT1 required longer incubation time (120 minutes). NAD+ (CHEBI:15846) Q96EB6 is required for deacetylation of Q04206 rela-sirt2-1 1 5 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey physical association physical association Fig. 3b tumor necrosis factor alpha (human) (TNF-alpha) (P01375) hist1h4a-sirt2 3 6 unspecified role unspecified role neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined nuclear localisation signal tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q8IXJ6-2:p.[Leu4Ala;Leu12Ala;His150Tyr] mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L A certain certain sequence position certain certain certain sequence position certain L A certain certain sequence position certain certain certain sequence position certain H Y Wed May 02 17:57:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu4ala,leu12ala,his150tyr' Tue Jun 30 16:10:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu4Ala;Leu12Ala;His150Tyr]' 7 unspecified role unspecified role neutral component neutral component possible acetyllys-17 naclys N-acetylated L-lysine NAcLys certain certain sequence position certain certain certain sequence position certain colocalization colocalization Suppl. Fig. S2a The Materials and Methods section states that the nuclear export signal (NES) of SIRT2 was disrupted by mutating K4 and K12 to alanines; however, the SIRT2 (isoform 2 with the NES at the N-terminus) contains leucine residues in these positions, which are a hallmark of NES motifs, hence these mutations are captured here as leucine-to-alanine mutations.