NTNU NTNU, Norwegian University of Science and Technology www.ntnu.no/home pastorcic-2003-1 Ets transcription factors ER81 and Elk1 regulate the transcription of the human presenilin 1 gene promoter. in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa autoradiography autoradiography Pastorcic M., Das H.K. Brain Res. Mol. Brain Res. (0169-328X) 2003 imex curation pastorcic-2003-2 Ets transcription factors ER81 and Elk1 regulate the transcription of the human presenilin 1 gene promoter. in vitro In vitro emsa supershift electrophoretic mobility supershift assay emsa supershift weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Pastorcic M., Das H.K. Brain Res. Mol. Brain Res. (0169-328X) 2003 imex curation pastorcic-2003-4 Ets transcription factors ER81 and Elk1 regulate the transcription of the human presenilin 1 gene promoter. yeast Saccharomyces cerevisiae Baker's yeast 1 hybrid one hybrid yeast one-hybrid yeast one hybrid one-hybrid 1 hybrid yeast one-hybrid one-hybrid yeast one hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Pastorcic M., Das H.K. Brain Res. Mol. Brain Res. (0169-328X) 2003 imex curation psen1_human_dna_probe presenilin 1 DNA probe, promoter ds dna double stranded deoxyribonucleic acid ds DNA ds dna human Homo sapiens Human CGCCAGAGCCGGAATGACGACAACGGT etv1_human ETS translocation variant 1 ETV1 ER81 Ets-related protein 81 protein protein human Homo sapiens Human MDGFYDQQVPYMVTNSQRGRNCNEKPTNVRKRKFINRDLAHDSEELFQDLSQLQETWLAEAQVPDNDEQFVPDYQAESLAFHGLPLKIKKEPHSPCSEISSACSQEQPFKFSYGEKCLYNVSAYDQKPQVGMRPSNPPTPSSTPVSPLHHASPNSTHTPKPDRAFPAHLPPSQSIPDSSYPMDHRFRRQLSEPCNSFPPLPTMPREGRPMYQRQMSEPNIPFPPQGFKQEYHDPVYEHNTMVGSAASQSFPPPLMIKQEPRDFAYDSEVPSCHSIYMRQEGFLAHPSRTEGCMFEKGPRQFYDDTCVVPEKFDGDIKQEPGMYREGPTYQRRGSLQLWQFLVALLDDPSNSHFIAWTGRGMEFKLIEPEEVARRWGIQKNRPAMNYDKLSRSLRYYYEKGIMQKVAGERYVYKFVCDPEALFSMAFPDNQRPLLKTDMERHINEEDTVPLSHFDESMAYMPEGGCCNPHPYNEGYVY elk1_human ETS domain-containing protein Elk-1 ELK1 protein protein human Homo sapiens Human MDPSVTLWQFLLQLLREQGNGHIISWTSRDGGEFKLVDAEEVARLWGLRKNKTNMNYDKLSRALRYYYDKNIIRKVSGQKFVYKFVSYPEVAGCSTEDCPPQPEVSVTSTMPNVAPAAIHAAPGDTVSGKPGTPKGAGMAGPGGLARSSRNEYMRSGLYSTFTIQSLQPQPPPHPRPAVVLPSAAPAGAAAPPSGSRSTSPSPLEACLEAEEAGLPLQVILTPPEAPNLKSEELNVEPGLGRALPPEVKVEGPKEELEVAGERGFVPETTKAEPEVPPQEGVPARLPAVVMDTAGQAGGHAASSPEISQPQKGRKPRDLELPLSPSLLGGPGPERTPGSGSGSGLQAPGPALTPSLLPTHTLTPVLLTPSSLPPSIHFWSTLSPIAPRSPAKLSFQFPSSGSAQVHIPSISVDGLSTPVVLSPGPQKP ets2_human Protein C-ets-2 ETS2 protein protein human Homo sapiens Human MNDFGIKNMDQVAPVANSYRGTLKRQPAFDTFDGSLFAVFPSLNEEQTLQEVPTGLDSISHDSANCELPLLTPCSKAVMSQALKATFSGFKKEQRRLGIPKNPWLWSEQQVCQWLLWATNEFSLVNVNLQRFGMNGQMLCNLGKERFLELAPDFVGDILWEHLEQMIKENQEKTEDQYEENSHLTSVPHWINSNTLGFGTEQAPYGMQTQNYPKGGLLDSMCPASTPSVLSSEQEFQMFPKSRLSSVSVTYCSVSQDFPGSNLNLLTNNSGTPKDHDSPENGADSFESSDSLLQSWNSQSSLLDVQRVPSFESFEDDCSQSLCLNKPTMSFKDYIQERSDPVEQGKPVIPAAVLAGFTGSGPIQLWQFLLELLSDKSCQSFISWTGDGWEFKLADPDEVARRWGKRKNKPKMNYEKLSRGLRYYYDKNIIHKTSGKRYVYRFVCDLQNLLGFTPEELHAILGVQPDTED psen1-etv1 1539638 n/a 1539641 autoradiography autoradiography unspecified role unspecified role bait bait radiolabel 32p radiolabel 32p radiolabel P32 32P 32P P32 autoradiography autoradiography undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false probe represents -22 to +6 of human PSEN1 promoter region g11t_g12t mutation mutation certain certain sequence position certain certain certain sequence position certain false ETV1_human (TF) did not bind to this mutated dsDNA probe n/a 1539642 unspecified role unspecified role prey prey in vitro In vitro direct interaction direct interaction false false false Fig 2 in vitro transcribed-translated (Promega) P50549 increases expression at RNA level of ENSG00000080815 ETV1 (P50549) increases expression at mRNA-level of PSEN1 (ENSG00000080815) as ascertained by reporter gene assay in SK-N-SH neuroblastoma cells, reported in Figure 5 psen1-elk1 1539638 n/a 1539641 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 false radiolabel 32p radiolabel 32p radiolabel P32 32P 32P P32 autoradiography autoradiography undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false g11t_g12t mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false n/a 1539643 unspecified role unspecified role prey prey direct interaction direct interaction false false false Fig4 P19419 decreases expression at RNA level of ENSG00000080815 ELK1 (P19419) decreases expression at mRNA level of PSEN1 (ENSG00000080815) as ascertained by reporter gene assay reported in Figure 6 psen1-etv1-1 1539639 n/a 1539641 unspecified role unspecified role bait bait radiolabel 32p radiolabel 32p radiolabel P32 32P 32P P32 autoradiography autoradiography undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false g11t_g12t mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false ETV1_human (TF) did not bind to this mutated dsDNA probe region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false probe represents -22 to +6 of human PSEN1 promoter region n/a 1539642 unspecified role unspecified role prey prey in vitro In vitro direct interaction direct interaction false false false Fig 2 in vitro transcribed-translated (Promega) ETV1 (P50549) increases expression at mRNA-level of PSEN1 (ENSG00000080815) as ascertained by reporter gene assay in SK-N-SH neuroblastoma cells, reported in Figure 5 P50549 increases expression at mRNA- evel of PSEN1 ENSG00000080815 psen1-elk1-2 1539639 n/a 1539641 unspecified role unspecified role bait bait radiolabel 32p radiolabel 32p radiolabel P32 32P 32P P32 autoradiography autoradiography undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false g11t_g12t mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false ETV1_human (TF) did not bind to this mutated dsDNA probe region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false probe represents -22 to +6 of human PSEN1 promoter region n/a 1539643 unspecified role unspecified role prey prey direct interaction direct interaction false false false Fig 4 in vitro transcribed-translated (Promega) P19419 decreases expression at RNA level of ENSG00000080815 ELK1 (P19419) decreases expression at mRNA level of PSEN1 (ENSG00000080815) as ascertained by reporter gene assay reported in Figure 6 psen1-etv1-2 1539640 n/a 1539641 unspecified role unspecified role bait bait n/a 1539642 unspecified role unspecified role prey prey association association false false false first paragraph Results psen1-ets2-2 1539640 n/a 1539641 unspecified role unspecified role bait bait n/a 1539644 unspecified role unspecified role prey prey association association false false false