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/ Homeodomain-Interacting Protein Kinase-2 (HIPK2) Phosphorylates HMGA1a at Ser-35, Thr-52, and Thr-77 and Modulates Its DNA Binding Affinity. Homeodomain-Interacting Protein Kinase-2 (HIPK2) Phosphorylates HMGA1a at Ser-35, Thr-52, and Thr-77 and Modulates Its DNA Binding Affinity. J. Proteome Res. (1535-3893) 2007 imex curation Zhang Q., Wang Y. Apoptosis - Interactions involving proteins with a function related to apoptosis Source of HIPK2 taken from manufacturer's website yinsheng.wang@ucr.edu yinsheng.wang@ucr.edu. 01-Feb-2008: Contacted by JYOTI. davet Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay fingerprinting peptide massfingerprinting fingerprinting Zhang Q., Wang Y. Apoptosis - Interactions involving proteins with a function related to apoptosis Source of HIPK2 taken from manufacturer's website yinsheng.wang@ucr.edu yinsheng.wang@ucr.edu Accepted 2008-FEB-10 by DAVET. 01-Feb-2008: Contacted by JYOTI. J. Proteome Res. (1535-3893) 2007 davet Only protein-protein interactions imex curation Homeodomain-Interacting Protein Kinase-2 (HIPK2) Phosphorylates HMGA1a at Ser-35, Thr-52, and Thr-77 and Modulates Its DNA Binding Affinity. Homeodomain-Interacting Protein Kinase-2 (HIPK2) Phosphorylates HMGA1a at Ser-35, Thr-52, and Thr-77 and Modulates Its DNA Binding Affinity. J. Proteome Res. (1535-3893) 2007 imex curation Zhang Q., Wang Y. Apoptosis - Interactions involving proteins with a function related to apoptosis Source of HIPK2 taken from manufacturer's website yinsheng.wang@ucr.edu yinsheng.wang@ucr.edu. 01-Feb-2008: Contacted by JYOTI. davet Only protein-protein interactions in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa predetermined predetermined participant predetermined Zhang Q., Wang Y. Apoptosis - Interactions involving proteins with a function related to apoptosis Source of HIPK2 taken from manufacturer's website yinsheng.wang@ucr.edu yinsheng.wang@ucr.edu Accepted 2008-FEB-10 by DAVET. 01-Feb-2008: Contacted by JYOTI. J. Proteome Res. (1535-3893) 2007 davet Only protein-protein interactions imex curation hipk2_human Homeodomain-interacting protein kinase 2 HIPK2 protein protein human Homo sapiens Human MAPVYEGMASHVQVFSPHTLQSSAFCSVKKLKIEPSSNWDMTGYGSHSKVYSQSKNIPLSQPATTTVSTSLPVPNPSLPYEQTIVFPGSTGHIVVTSASSTSVTGQVLGGPHNLMRRSTVSLLDTYQKCGLKRKSEEIENTSSVQIIEEHPPMIQNNASGATVATATTSTATSKNSGSNSEGDYQLVQHEVLCSMTNTYEVLEFLGRGTFGQVVKCWKRGTNEIVAIKILKNHPSYARQGQIEVSILARLSTESADDYNFVRAYECFQHKNHTCLVFEMLEQNLYDFLKQNKFSPLPLKYIRPVLQQVATALMKLKSLGLIHADLKPENIMLVDPSRQPYRVKVIDFGSASHVSKAVCSTYLQSRYYRAPEIILGLPFCEAIDMWSLGCVIAELFLGWPLYPGASEYDQIRYISQTQGLPAEYLLSAGTKTTRFFNRDTDSPYPLWRLKTPDDHEAETGIKSKEARKYIFNCLDDMAQVNMTTDLEGSDMLVEKADRREFIDLLKKMLTIDADKRITPIETLNHPFVTMTHLLDFPHSTHVKSCFQNMEICKRRVNMYDTVNQSKTPFITHVAPSTSTNLTMTFNNQLTTVHNQAPSSTSATISLANPEVSILNYPSTLYQPSAASMAAVAQRSMPLQTGTAQICARPDPFQQALIVCPPGFQGLQASPSKHAGYSVRMENAVPIVTQAPGAQPLQIQPGLLAQQAWPSGTQQILLPPAWQQLTGVATHTSVQHATVIPETMAGTQQLADWRNTHAHGSHYNPIMQQPALLTGHVTLPAAQPLNVGVAHVMRQQPTSTTSSRKSKQHQSSVRNVSTCEVSSSQAISSPQRSKRVKENTPPRCAMVHSSPACSTSVTCGWGDVASSTTRERQRQTIVIPDTPSPTVSVITISSDTDEEEEQKHAPTSTVSKQRKNVISCVTVHDSPYSDSSSNTSPYSVQQRAGHNNANAFDTKGSLENHCTGNPRTIIVPPLKTQASEVLVECDSLVPVNTSHHSSSYKSKSSSNVTSTSGHSSGSSSGAITYRQQRPGPHFQQQQPLNLSQAQQHITTDRTGSHRRQQAYITPTMAQAPYSFPHNSPSHGTVHPHLAAAAAAAHLPTQPHLYTYTAPAALGSTGTVAHLVASQGSARHTVQHTAYPASIVHQVPVSMGPRVLPSPTIHPSQYPAQFAHQTYISASPASTVYTGYPLSPAKVNQYPYI DIP protein Q9H2X6 original sequence version: 132 6022D5710E8D2D93 germ_line_epsilon Human germ line epsilon promoter oligonucleotide ds dna double stranded deoxyribonucleic acid ds DNA ds dna human Homo sapiens Human CCACTGCCCGGCACAGAAATAACAACCACGGTTACT 4FF3D22FAC7750CB cdk1_human Cyclin-dependent kinase 1 p34 protein kinase Cell division protein kinase 1 CDK1 CDC2 Cell division control protein 2 homolog CDC28A CDKN1 P34CDC2 protein protein human Homo sapiens Human MEDYTKIEKIGEGTYGVVYKGRHKTTGQVVAMKKIRLESEEEGVPSTAIREISLLKELRHPNIVSLQDVLMQDSRLYLIFEFLSMDLKKYLDSIPPGQYMDSSLVKSYLYQILQGIVFCHSRRVLHRDLKPQNLLIDDKGTIKLADFGLARAFGIPIRVYTHEVVTLWYRSPEVLLGSARYSTPVDIWSIGTIFAELATKKPLFHGDSEIDQLFRIFRALGTPNNEVWPEVESLQDYKNTFPKWKPGSLASHVKNLDENGLDLLSKMLIYDPAKRISGKMALNHPYFNDLDNQIKKM DIP protein P06493 original sequence version: 185 DIP protein P06493 original sequence version: 162 DIP protein P06493 original sequence version: 184 DIP protein P06493 original sequence version: 97 DIP protein P06493 original sequence version: 192 942D79448EFE490A p17096-1 High mobility group protein HMG-I/HMG-Y HMGA1a HMGA1 HMGIY High mobility group AT-hook protein 1 High mobility group protein R protein protein human Homo sapiens Human MSESSSKSSQPLASKQEKDGTEKRGRGRPRKQPPVSPGTALVGSQKEPSEVPTPKRPRGRPKGSKNKGAAKTRKTTTTPGRKPRGRPKKLEKEEEEGISQESSEEEQ E9C4E3F2200914B8 p17096-2 High mobility group protein HMG-I/HMG-Y HMGA1b HMGA1 HMGIY High mobility group AT-hook protein 1 High mobility group protein R protein protein human Homo sapiens Human MSESSSKSSQPLASKQEKDGTEKRGRGRPRKQPPKEPSEVPTPKRPRGRPKGSKNKGAAKTRKTTTTPGRKPRGRPKKLEKEEEEGISQESSEEEQ Contains a phosphothreonine at position 67. Contains a phosphothreonine at position 42. B82DCAA29E6D18FD hipk2-hmga1-1 1 6 enzyme target enzyme target substrate substrate neutral component neutral component ecobb Escherichia coli (strain B / BL21) 3 enzyme enzyme neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis 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. phosphorylation phosphorylation reaction phosphorylation Here, we found that HIPK2 phosphorylates HMGA1a at Ser-35, Thr-52, and Thr-77, and HMGA1b at Thr-41 and Thr-66. Phosphorylation of Ser-36, Thr-53 and Thr-78 of P17096-1 Fig. 1 hipk2-hmga1-2 1 7 enzyme target enzyme target substrate substrate neutral component neutral component ecobb Escherichia coli (strain B / BL21) 3 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-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. phosphorylation phosphorylation reaction phosphorylation Here, we found that HIPK2 phosphorylates HMGA1a at Ser-35, Thr-52, and Thr-77, and HMGA1b at Thr-41 and Thr-66. Phosphorylation of Thr-42 and Thr-67 of P17096-2 Supp. Fig. 1 cdc2-hmga1-1 1 5 enzyme enzyme neutral component neutral component 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. 6 enzyme target enzyme target substrate substrate neutral component neutral component ecobb Escherichia coli (strain B / BL21) phosphorylation phosphorylation reaction phosphorylation In addition, we demonstrated that cdc2, which is known to phosphorylate HMGA1 proteins, could induce the phosphorylation of HMGA1 proteins at the same Ser/Thr sites. Phosphorylation of Ser-36, Thr-53 and Thr-77 of P17096-1 Fig. 2 hmga1-dna-1 2 4 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Moreover, we found that the HIPK2-phosphorylated HMGA1a reduced the binding affinity of HMGA1a to human germ line epsilon promoter Fig. 3A, Fig 4 Phosphorylation of HMGA1a by HIPK2 reduced the binding affinity of HMGA1a to the human germ line epsilon promote. A more profound drop was induced by cdc2 phosphorylation which preferentially phosphorylates Thr-53 rather than Thr-77 preferred by HIPK2