NTNU NTNU, Norwegian University of Science and Technology www.ntnu.no/home yanagihara-2005-2 Paired-like homeoprotein ESXR1 acts as a sequence-specific transcriptional repressor of the human K-ras gene. in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa autoradiography autoradiography Yanagihara M., Ishikawa S., Naito M., Nakajima J., Aburatani H., Hatakeyama M. human origin of homeoprotein EZXR1 was ascertained by checking the reference Ozawa H et al, 2004, PMID:15235584 (given in the paper as source for EZXR1) mhata@igm.hokudai.ac.jp Oncogene (0950-9232) 2005 imex curation yanagihara-2005-1 Paired-like homeoprotein ESXR1 acts as a sequence-specific transcriptional repressor of the human K-ras gene. human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Yanagihara M., Ishikawa S., Naito M., Nakajima J., Aburatani H., Hatakeyama M. human origin of homeoprotein EZXR1 was ascertained by checking the reference Ozawa H et al, 2004, PMID:15235584 (given in the paper as source for EZXR1) mhata@igm.hokudai.ac.jp Oncogene (0950-9232) 2005 imex curation yanagihara-2005-3 Paired-like homeoprotein ESXR1 acts as a sequence-specific transcriptional repressor of the human K-ras gene. in vitro In vitro emsa supershift electrophoretic mobility supershift assay emsa supershift autoradiography autoradiography Yanagihara M., Ishikawa S., Naito M., Nakajima J., Aburatani H., Hatakeyama M. human origin of homeoprotein EZXR1 was ascertained by checking the reference Ozawa H et al, 2004, PMID:15235584 (given in the paper as source for EZXR1) mhata@igm.hokudai.ac.jp Oncogene (0950-9232) 2005 imex curation kras_human_dna_51bp_intronic K-ras intronic regulatory region, 51bp fragment dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid chemical synthesis Chemical synthesis (Chemical synthesis) TAAAAGGGTAAAGGACATCTTAATGTTATTATGAAAACAGTTTTGACCTCT esx1_human Homeobox protein ESX1 ESX1 ESX1L ESX1R Extraembryonic, spermatogenesis, homeobox 1 protein protein human Homo sapiens Human MESLRGYTHSDIGYRSLAVGEDIEEVNDEKLTVTSLMARGGEDEENTRSKPEYGTEAENNVGTEGSVPSDDQDREGGGGHEPEQQQEEPPLTKPEQQQEEPPLLELKQEQEEPPQTTVEGPQPAEGPQTAEGPQPPERKRRRRTAFTQFQLQELENFFDESQYPDVVARERLAARLNLTEDRVQVWFQNRRAKWKRNQRVLMLRNTATADLAHPLDMFLGGAYYAAPALDPALCVHLVPQLPRPPVLPVPPMPPRPPMVPMPPRPPIAPMPPMAPVPPGSRMAPVPPGPRMAPVPPWPPMAPVPPWPPMAPVPTGPPMAPVPPGPPMARVPPGPPMARVPPGPPMAPLPPGPPMAPLPPGPPMAPLPPGPPMAPLPPRSHVPHTGLAPVHITWAPVINSYYACPFF kras_human_gene Kirsten rat sarcoma viral oncogene homolog gene gene human Homo sapiens Human kras-esx1-2 1444935 n/a 1444938 unspecified role unspecified role bait bait region 32p radiolabel 32p radiolabel P32 32P 32P P32 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 chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1444938 competitor competitor competitor prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false a23c_t29g_t30g_a31c mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1444939 unspecified role unspecified role prey prey region sufficient for dna binding and transcription regulation sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 physical association physical association false false false Fig 4c kras-esx1-1 1444935 n/a 1444938 unspecified role unspecified role bait bait region 32p radiolabel 32p radiolabel P32 32P 32P P32 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 chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1444939 unspecified role unspecified role prey prey region sufficient for dna binding and transcription regulation sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false mutation is deletion of C-terminus containing a repeat region and leaving the N-terminal amino acids 1-229 encompassing e.g. the homeodomain intact (cloning of deletion mutant described in PMID:15235584) chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 physical association physical association false false false Fig 4c esx1-kras 1444936 n/a K-ras 1444940 unspecified role unspecified role prey prey n/a esxr1 1444939 unspecified role unspecified role bait bait region sufficient for dna binding and transcription regulation sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false mutation is deletion of C-terminus containing a repeat region and leaving the N-terminal amino acids 1-229 encompassing e.g. the homeodomain intact (cloning of deletion mutant described in PMID:15235584) region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference association association false false false Fig 4d Q8N693 decreases expression at RNA level of ENSG00000133703 Q8N693 decreases expression at RNA level of ENSG00000133703 (decreases activity of intronic regulatory region (SO:0001492) as measured in reporter gene assay). Reported in Figure 4b kras-esx1-1 1444937 n/a 1444938 unspecified role unspecified role bait bait region 32p radiolabel 32p radiolabel P32 32P 32P P32 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 chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1444939 unspecified role unspecified role prey prey region sufficient for dna binding and transcription regulation sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false mutation is deletion of C-terminus containing a repeat region and leaving the N-terminal amino acids 1-229 encompassing e.g. the homeodomain intact (cloning of deletion mutant described in PMID:15235584) chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 physical association physical association false false false Fig 4c