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/ The basis for IL-2-induced IL-2 receptor alpha chain gene regulation: importance of two widely separated IL-2 response elements. The basis for IL-2-induced IL-2 receptor alpha chain gene regulation: importance of two widely separated IL-2 response elements. Immunity (1074-7613) 2001 imex curation Kim H.P., Kelly J., Leonard W.J. IBD - Inflammatory bowel disease Recombinant human HMG-I and Stat5a are described in PMID:8407950 and PMID:10022878, respectively. wjl@helix.nih.gov 26-Feb-2015: Contacted by IntAct-Help. Only protein-protein interactions human-yt Homo sapiens yt Human T/NK cell leukemia ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Kim H.P., Kelly J., Leonard W.J. IBD - Inflammatory bowel disease Recombinant human HMG-I and Stat5a are described in PMID:8407950 and PMID:10022878, respectively. wjl@helix.nih.gov Accepted 2015-JAN-29 by ORCHARD Immunity (1074-7613) 2001 Only protein-protein interactions imex curation The basis for IL-2-induced IL-2 receptor alpha chain gene regulation: importance of two widely separated IL-2 response elements. The basis for IL-2-induced IL-2 receptor alpha chain gene regulation: importance of two widely separated IL-2 response elements. Immunity (1074-7613) 2001 imex curation Kim H.P., Kelly J., Leonard W.J. IBD - Inflammatory bowel disease Recombinant human HMG-I and Stat5a are described in PMID:8407950 and PMID:10022878, respectively. wjl@helix.nih.gov 26-Feb-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro uracil interference assay uracil interference assay autoradiography autoradiography Kim H.P., Kelly J., Leonard W.J. IBD - Inflammatory bowel disease Recombinant human HMG-I and Stat5a are described in PMID:8407950 and PMID:10022878, respectively. wjl@helix.nih.gov Accepted 2015-JAN-29 by ORCHARD Immunity (1074-7613) 2001 Only protein-protein interactions imex curation The basis for IL-2-induced IL-2 receptor alpha chain gene regulation: importance of two widely separated IL-2 response elements. The basis for IL-2-induced IL-2 receptor alpha chain gene regulation: importance of two widely separated IL-2 response elements. Immunity (1074-7613) 2001 imex curation Kim H.P., Kelly J., Leonard W.J. IBD - Inflammatory bowel disease Recombinant human HMG-I and Stat5a are described in PMID:8407950 and PMID:10022878, respectively. wjl@helix.nih.gov 26-Feb-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro dnase 1 footprinting DNase I footprinting dnase 1 footprinting autoradiography autoradiography Kim H.P., Kelly J., Leonard W.J. IBD - Inflammatory bowel disease Recombinant human HMG-I and Stat5a are described in PMID:8407950 and PMID:10022878, respectively. wjl@helix.nih.gov Accepted 2015-JAN-29 by ORCHARD Immunity (1074-7613) 2001 Only protein-protein interactions imex curation The basis for IL-2-induced IL-2 receptor alpha chain gene regulation: importance of two widely separated IL-2 response elements. The basis for IL-2-induced IL-2 receptor alpha chain gene regulation: importance of two widely separated IL-2 response elements. Immunity (1074-7613) 2001 imex curation Kim H.P., Kelly J., Leonard W.J. IBD - Inflammatory bowel disease Recombinant human HMG-I and Stat5a are described in PMID:8407950 and PMID:10022878, respectively. wjl@helix.nih.gov 26-Feb-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa autoradiography autoradiography Kim H.P., Kelly J., Leonard W.J. IBD - Inflammatory bowel disease Recombinant human HMG-I and Stat5a are described in PMID:8407950 and PMID:10022878, respectively. wjl@helix.nih.gov Accepted 2015-JAN-29 by ORCHARD Immunity (1074-7613) 2001 Only protein-protein interactions imex curation The basis for IL-2-induced IL-2 receptor alpha chain gene regulation: importance of two widely separated IL-2 response elements. The basis for IL-2-induced IL-2 receptor alpha chain gene regulation: importance of two widely separated IL-2 response elements. Immunity (1074-7613) 2001 imex curation Kim H.P., Kelly J., Leonard W.J. IBD - Inflammatory bowel disease Recombinant human HMG-I and Stat5a are described in PMID:8407950 and PMID:10022878, respectively. wjl@helix.nih.gov 26-Feb-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro emsa supershift electrophoretic mobility supershift assay emsa supershift autoradiography autoradiography Kim H.P., Kelly J., Leonard W.J. IBD - Inflammatory bowel disease Recombinant human HMG-I and Stat5a are described in PMID:8407950 and PMID:10022878, respectively. wjl@helix.nih.gov Accepted 2015-JAN-29 by ORCHARD Immunity (1074-7613) 2001 Only protein-protein interactions imex curation il2ra_human_gas1-site_dna-probe Human IL2RA gene promoter gamma-activated sequence-1 DNA probe dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid human Homo sapiens Human GAAGGCACCTTTCTCAGAACTGTGGAAATTC sta5a_human Signal transducer and activator of transcription 5A STAT5A STAT5 protein protein human Homo sapiens Human MAGWIQAQQLQGDALRQMQVLYGQHFPIEVRHYLAQWIESQPWDAIDLDNPQDRAQATQLLEGLVQELQKKAEHQVGEDGFLLKIKLGHYATQLQKTYDRCPLELVRCIRHILYNEQRLVREANNCSSPAGILVDAMSQKHLQINQTFEELRLVTQDTENELKKLQQTQEYFIIQYQESLRIQAQFAQLAQLSPQERLSRETALQQKQVSLEAWLQREAQTLQQYRVELAEKHQKTLQLLRKQQTIILDDELIQWKRRQQLAGNGGPPEGSLDVLQSWCEKLAEIIWQNRQQIRRAEHLCQQLPIPGPVEEMLAEVNATITDIISALVTSTFIIEKQPPQVLKTQTKFAATVRLLVGGKLNVHMNPPQVKATIISEQQAKSLLKNENTRNECSGEILNNCCVMEYHQATGTLSAHFRNMSLKRIKRADRRGAESVTEEKFTVLFESQFSVGSNELVFQVKTLSLPVVVIVHGSQDHNATATVLWDNAFAEPGRVPFAVPDKVLWPQLCEALNMKFKAEVQSNRGLTKENLVFLAQKLFNNSSSHLEDYSGLSVSWSQFNRENLPGWNYTFWQWFDGVMEVLKKHHKPHWNDGAILGFVNKQQAHDLLINKPDGTFLLRFSDSEIGGITIAWKFDSPERNLWNLKPFTTRDFSIRSLADRLGDLSYLIYVFPDRPKDEVFSKYYTPVLAKAVDGYVKPQIKQVVPEFVNASADAGGSSATYMDQAPSPAVCPQAPYNMYPQNPDHVLDQDGEFDLDETMDVARHVEELLRRPMDSLDSRLSPPAGLFTSARGSLS DIP protein P42229 original sequence version: 162 C64237295F88CFBE il2ra_human_gas2-site_dna-probe Human IL2RA gene promoter gamma-activated sequence-2 DNA probe dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid human Homo sapiens Human CTTTCTTTCATTCTAGGAATTCTGTGATCAA sta5b_human Signal transducer and activator of transcription 5B STAT5B protein protein human Homo sapiens Human MAVWIQAQQLQGEALHQMQALYGQHFPIEVRHYLSQWIESQAWDSVDLDNPQENIKATQLLEGLVQELQKKAEHQVGEDGFLLKIKLGHYATQLQNTYDRCPMELVRCIRHILYNEQRLVREANNGSSPAGSLADAMSQKHLQINQTFEELRLVTQDTENELKKLQQTQEYFIIQYQESLRIQAQFGPLAQLSPQERLSRETALQQKQVSLEAWLQREAQTLQQYRVELAEKHQKTLQLLRKQQTIILDDELIQWKRRQQLAGNGGPPEGSLDVLQSWCEKLAEIIWQNRQQIRRAEHLCQQLPIPGPVEEMLAEVNATITDIISALVTSTFIIEKQPPQVLKTQTKFAATVRLLVGGKLNVHMNPPQVKATIISEQQAKSLLKNENTRNDYSGEILNNCCVMEYHQATGTLSAHFRNMSLKRIKRSDRRGAESVTEEKFTILFESQFSVGGNELVFQVKTLSLPVVVIVHGSQDNNATATVLWDNAFAEPGRVPFAVPDKVLWPQLCEALNMKFKAEVQSNRGLTKENLVFLAQKLFNNSSSHLEDYSGLSVSWSQFNRENLPGRNYTFWQWFDGVMEVLKKHLKPHWNDGAILGFVNKQQAHDLLINKPDGTFLLRFSDSEIGGITIAWKFDSQERMFWNLMPFTTRDFSIRSLADRLGDLNYLIYVFPDRPKDEVYSKYYTPVPCESATAKAVDGYVKPQIKQVVPEFVNASADAGGGSATYMDQAPSPAVCPQAHYNMYPQNPDSVLDTDGDFDLEDTMDVARRVEELLGRPMDSQWIPHAQS AA2F1CAB20955ACA il2ra_human_gene Human IL2RA (CD25) gene gene gene human Homo sapiens Human il2ra_human_+3317/+3478_dna-probe Human IL2RA gene promoter region +3317 to +3478 DNA probe dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid human Homo sapiens Human CAACAGATGCAATTTCTAGCTGATTCATAAGGGCAGATTTTCATTCTGAAAATTTCCAAAGCCACCATTTAACATGGCAAGGGTTTATGAGGACATATTAGTCAACTTGAGTGTTTTTAGTCTCTATATTACTGGGCTCTGGGATGTGGTCAGAAAGAGAAT hmga1_human High mobility group protein HMG-I/HMG-Y High mobility group AT-hook protein 1 HMGA1 HMGIY High mobility group protein R protein protein human Homo sapiens Human MSESSSKSSQPLASKQEKDGTEKRGRGRPRKQPPVSPGTALVGSQKEPSEVPTPKRPRGRPKGSKNKGAAKTRKTTTTPGRKPRGRPKKLEKEEEEGISQESSEEEQ DIP protein P17096 original sequence version: 100 E9C4E3F2200914B8 stat5a-il2ra-1 1 10 unspecified role unspecified role prey prey promoter region prr3 binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined intron 1 region prr4 binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role bait bait association association Fig. 8a human interleukin-2 (IL-2) (P60568) P60568 increases binding of P42229 to ENSG00000134460 P42229 increases expression at RNA level of ENSG00000134460 P60568 indirectly increases expression at RNA level of ENSG00000134460 stat5b-il2ra-1 1 9 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey promoter region prr3 binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined intron 1 region prr4 binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig. 8a human interleukin-2 (IL-2) (P60568) P60568 increases binding of P51692 to ENSG00000134460 P51692 increases expression at RNA level of ENSG00000134460 hmga1-il2ra-1 1 12 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey promoter region prr3 binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined intron 1 region prr4 binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig. 8a human interleukin-2 (IL-2) (P60568) P60568 increases binding of P17096 to ENSG00000134460 hmga1-il2ra-2 1 12 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey promoter region prr1/2 binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig. 8a il2ra-stat5a-3 5 6 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 chemical synthesis Chemical synthesis (Chemical synthesis) 7 unspecified role unspecified role prey prey human-yt Homo sapiens yt Human T/NK cell leukemia physical association physical association Fig. 5c human interleukin-2 (IL-2) (P60568) il2ra-stat5b-2 5 9 unspecified role unspecified role prey prey human-yt Homo sapiens yt Human T/NK cell leukemia 6 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 chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association Fig. 5c human interleukin-2 (IL-2) (P60568) il2ra-stat5b-1 5 8 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 chemical synthesis Chemical synthesis (Chemical synthesis) 9 unspecified role unspecified role prey prey human-yt Homo sapiens yt Human T/NK cell leukemia physical association physical association Fig. 5c human interleukin-2 (IL-2) (P60568) il2ra-stat5a-1 5 7 unspecified role unspecified role prey prey human-yt Homo sapiens yt Human T/NK cell leukemia 8 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 chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association Fig. 5c human interleukin-2 (IL-2) (P60568) il2ra-hmga1-2 4 11 unspecified role unspecified role bait bait t116g_t117g_t127g_a128g mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain t39g_t40g_a51g_a52g_t69g_t70g_t86g_a87g_a97g_t98g_t116g_t117g_t127g_a128g mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain t69g_t70g_t86g_a87g_a97g_t98g mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined t69g_t70g_t86g_a87g_a97g_t98g_t116g_t117g_t127g_a128g mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain t39g_t40g_a51g_a52g mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain t39g_t40g_a51g_a52g_t116g_t117g_t127g_a128g mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain t39g_t40g_a51g_a52g_t69g_t70g_t86g_a87g_a97g_t98g mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 12 unspecified role unspecified role prey prey direct interaction direct interaction Fig. 7e Labeled DNA probe was incubated with purified recombinant HMG-I protein; however, the expression system was not specified. The sequence of the probe was derived from the positions given in the figure legend of Fig. 7E and the sequence of the human IL2RA gene shown in Fig. 2B. il2ra-hmga1-1 3 10 unspecified role unspecified role bait bait region radiolabelled radiolabel radiolabeled radiolabelled undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined intron 1 region prr4 sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 12 unspecified role unspecified role prey prey direct interaction direct interaction Fig. 7a, 7b, 7c The labeled DNA fragment used in this interaction corresponds to the IL2RA gene region ranging from +3269 to +3596, located in the first intron. Labeled DNA was incubated with purified recombinant HMG-I protein; however, the expression system was not specified. il2ra-stat5a-2 2 7 unspecified role unspecified role prey prey trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 10 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 intron 1 region prr4 sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction Fig. 5d The labeled DNA fragment used in this interaction corresponds to the IL2RA gene region ranging from +3474 to +3622, located in the first intron. Uracil interference identified two canonical and one non-canonical gamma-interferon activated sequence motifs in the first intron of the IL-3 gene that bound STAT5A, with the canonical motifs showing higher affinity for STAT5A. In the canonical motifs, thymines in position 1 and 9 showed strong affinity for STAT5A, while those in positions 2 and 8 showed weaker binding. Thymines in positions 1 and 8 of the non-canonical motif also showed weaker binding to STAT5A.