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 Suppression of the HSF1-mediated proteotoxic stress response by the metabolic stress sensor AMPK. Suppression of the HSF1-mediated proteotoxic stress response by the metabolic stress sensor AMPK. EMBO J. (0261-4189) 2015 imex curation Dai S., Tang Z., Cao J., Zhou W., Li H., Sampson S., Dai C. Chengkai.Dai@jax.org 26-Feb-2015: Contacted by IntAct-Help. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Dai S., Tang Z., Cao J., Zhou W., Li H., Sampson S., Dai C. Chengkai.Dai@jax.org Accepted 2015-FEB-23 by MARTA EMBO J. (0261-4189) 2015 Only protein-protein interactions imex curation Suppression of the HSF1-mediated proteotoxic stress response by the metabolic stress sensor AMPK. Suppression of the HSF1-mediated proteotoxic stress response by the metabolic stress sensor AMPK. EMBO J. (0261-4189) 2015 imex curation Dai S., Tang Z., Cao J., Zhou W., Li H., Sampson S., Dai C. Chengkai.Dai@jax.org 26-Feb-2015: Contacted by IntAct-Help. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. density sedimentation cosedimentation through density gradient density sedimentation western blot western blot Immuno blot Immuno blot Dai S., Tang Z., Cao J., Zhou W., Li H., Sampson S., Dai C. Chengkai.Dai@jax.org Accepted 2015-FEB-23 by MARTA EMBO J. (0261-4189) 2015 Only protein-protein interactions imex curation lmna_human Prelamin-A/C LMNA LMN1 protein protein human Homo sapiens Human METPSQRRATRSGAQASSTPLSPTRITRLQEKEDLQELNDRLAVYIDRVRSLETENAGLRLRITESEEVVSREVSGIKAAYEAELGDARKTLDSVAKERARLQLELSKVREEFKELKARNTKKEGDLIAAQARLKDLEALLNSKEAALSTALSEKRTLEGELHDLRGQVAKLEAALGEAKKQLQDEMLRRVDAENRLQTMKEELDFQKNIYSEELRETKRRHETRLVEIDNGKQREFESRLADALQELRAQHEDQVEQYKKELEKTYSAKLDNARQSAERNSNLVGAAHEELQQSRIRIDSLSAQLSQLQKQLAAKEAKLRDLEDSLARERDTSRRLLAEKEREMAEMRARMQQQLDEYQELLDIKLALDMEIHAYRKLLEGEEERLRLSPSPTSQRSRGRASSHSSQTQGGGSVTKKRKLESTESRSSFSQHARTSGRVAVEEVDEEGKFVRLRNKSNEDQSMGNWQIKRQNGDDPLLTYRFPPKFTLKAGQVVTIWAAGAGATHSPPTDLVWKAQNTWGCGNSLRTALINSTGEEVAMRKLVRSVTVVEDDEDEDGDDLLHHHHGSHCSSSGDPAEYNLRSRTVLCGTCGQPADKASASGSGAQVGGPISSGSSASSVTVTRSYRSVGGSGGGSFGDNLVTRSYLLGNSSPRTQSPQNCSIM E0855F7699F0318B hsf1_human Heat shock factor protein 1 Heat shock transcription factor 1 HSF1 HSTF1 protein protein human Homo sapiens Human MDLPVGPGAAGPSNVPAFLTKLWTLVSDPDTDALICWSPSGNSFHVFDQGQFAKEVLPKYFKHNNMASFVRQLNMYGFRKVVHIEQGGLVKPERDDTEFQHPCFLRGQEQLLENIKRKVTSVSTLKSEDIKIRQDSVTKLLTDVQLMKGKQECMDSKLLAMKHENEALWREVASLRQKHAQQQKVVNKLIQFLISLVQSNRILGVKRKIPLMLNDSGSAHSMPKYSRQFSLEHVHGSGPYSAPSPAYSSSSLYAPDAVASSGPIISDITELAPASPMASPGGSIDERPLSSSPLVRVKEEPPSPPQSPRVEEASPGRPSSVDTLLSPTALIDSILRESEPAPASVTALTDARGHTDTEGRPPSPPPTSTPEKCLSVACLDKNELSDHLDAMDSNLDNLQTMLSSHGFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPRPPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAEDPTISLLTGSEPPKAKDPTVS DIP protein Q00613 original sequence version: 97 DIP protein Q00613 original sequence version: 157 DIP protein Q00613 original sequence version: 161 DIP protein Q00613-1 original sequence version: 161 735074507C954365 aapk1_rat 5'-AMP-activated protein kinase catalytic subunit alpha-1 Prkaa1 Ampk1 Hydroxymethylglutaryl-CoA reductase kinase Tau-protein kinase PRKAA1 Acetyl-CoA carboxylase kinase protein protein rat Rattus norvegicus (Rat) Rat MRRLSSWRKMATAEKQKHDGRVKIGHYILGDTLGVGTFGKVKVGKHELTGHKVAVKILNRQKIRSLDVVGKIRREIQNLKLFRHPHIIKLYQVISTPSDIFMVMEYVSGGELFDYICKNGRLDEKESRRLFQQILSGVDYCHRHMVVHRDLKPENVLLDAHMNAKIADFGLSNMMSDGEFLRTSCGSPNYAAPEVISGRLYAGPEVDIWSSGVILYALLCGTLPFDDDHVPTLFKKICDGIFYTPQYLNPSVISLLKHMLQVDPMKRATIKDIREHEWFKQDLPKYLFPEDPSYSSTMIDDEALKEVCEKFECSEEEVLSCLYNRNHQDPLAVAYHLIIDNRRIMNEAKDFYLATSPPDSFLDDHHLTRPHPERVPFLVAETPRARHTLDELNPQKSKHQGVRKAKWHLGIRSQSRPNDIMAEVCRAIKQLDYEWKVVNPYYLRVRRKNPVTSTFSKMSLQLYQVDSRTYLLDFRSIDDEITEAKSGTATPQRSGSISNYRSCQRSDSDAEAQGKPSEVSLTSSVTSLDSSPVDVAPRPGSHTIEFFEMCANLIKILAQ DIP protein P54645 original sequence version: 125 DIP protein P54645 original sequence version: 104 DIP protein P54645 original sequence version: 138 DIP protein P54645 original sequence version: 118 Responsible for the regulation of fatty acid synthesis by phosphorylation of acetyl-CoA carboxylase. It also regulates cholesterol synthesis via phosphorylation and inactivation of hormone-sensitive lipase and hydroxymethylglutaryl-CoA reductase. Appears to act as a metabolic stress-sensing protein kinase switching off biosynthetic pathways when cellular ATP levels are depleted and when 5'-AMP rises in response to fuel limitation and/or hypoxia. This is a catalytic subunit mint A869C340A85785ED hsf1-prkaa1-1 1 5 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell 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 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell 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 physical association physical association sf2f hsf1-prkaa1-2 2 3 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level 4 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component subcellular prep subcellular preparation subcellular prep over-expressed over expressed level over-expressed 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 true colocalization colocalization f3i f3j Despite no impacts on HSF1 expression, AMPKaCA impaired constitutive nuclear translocation of HSF1WT in HEK293T cells, indicated by diminished nuclear–cytoplasmic ratio of HSF1 levels (Fig 3I). S121A mutation not only enhanced HSF1 nuclear translocation under basal conditions but also rendered HSF1 resistant to the effect of AMPKaCA (Fig 3I). Similarly, while metformin impaired HSF1WT nuclear translocation, it exerted little effect on HSF1S121A mutants (Fig 3J). Thus, this defect in nuclear translocation due to Ser121 phosphorylation at least in part contributes to reduced HSF1–DNA binding hsf1-lmna 2 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level 3 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level colocalization colocalization f1j Next, we investigated whether metformin affected HSF1 nuclear translocation, a prerequisite for its DNA binding. As expected, heat shock caused most HSF1 proteins to translocate from the cytoplasm to the nucleus (Fig 1J). Importantly, metformin impeded this translocation (Fig 1J). Thus, metformin impairs HSF1–DNA binding at least in part through blockade of its nuclear translocation.