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/ TOUSLED kinase activity oscillates during the cell cycle and interacts with chromatin regulators. TOUSLED kinase activity oscillates during the cell cycle and interacts with chromatin regulators. Plant Physiol. (0032-0889) 2004 imex curation Ehsan H., Reichheld JP., Durfee T., Roe JL. Chromatin - Epigenetic interactions resulting in chromatin modulation jroe@ksu.edu 01-Sep-2009: Contacted by JYOTI. Arabidopsis cDNA library made from aerial parts of the palnt including flowers. orchard Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay autoradiography autoradiography Ehsan H., Reichheld JP., Durfee T., Roe JL. Chromatin - Epigenetic interactions resulting in chromatin modulation The source organism of the MBP or Histone 3 proteins could not be established and interactions involving this proteins were therefore not curated. jroe@ksu.edu Accepted 2009-AUG-24 by ORCHARD. 01-Sep-2009: Contacted by JYOTI. Arabidopsis cDNA library made from aerial parts of the palnt including flowers. Plant Physiol. (0032-0889) 2004 orchard Only protein-protein interactions imex curation TOUSLED kinase activity oscillates during the cell cycle and interacts with chromatin regulators. TOUSLED kinase activity oscillates during the cell cycle and interacts with chromatin regulators. Plant Physiol. (0032-0889) 2004 imex curation Ehsan H., Reichheld JP., Durfee T., Roe JL. Chromatin - Epigenetic interactions resulting in chromatin modulation jroe@ksu.edu 01-Sep-2009: Contacted by JYOTI. Arabidopsis cDNA library made from aerial parts of the palnt including flowers. orchard Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Ehsan H., Reichheld JP., Durfee T., Roe JL. Chromatin - Epigenetic interactions resulting in chromatin modulation jroe@ksu.edu Accepted 2009-AUG-24 by ORCHARD. 01-Sep-2009: Contacted by JYOTI. Arabidopsis cDNA library made from aerial parts of the palnt including flowers. Plant Physiol. (0032-0889) 2004 orchard Only protein-protein interactions imex curation TOUSLED kinase activity oscillates during the cell cycle and interacts with chromatin regulators. TOUSLED kinase activity oscillates during the cell cycle and interacts with chromatin regulators. Plant Physiol. (0032-0889) 2004 imex curation Ehsan H., Reichheld JP., Durfee T., Roe JL. Chromatin - Epigenetic interactions resulting in chromatin modulation jroe@ksu.edu 01-Sep-2009: Contacted by JYOTI. Arabidopsis cDNA library made from aerial parts of the palnt including flowers. orchard Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Ehsan H., Reichheld JP., Durfee T., Roe JL. Chromatin - Epigenetic interactions resulting in chromatin modulation jroe@ksu.edu Accepted 2009-AUG-24 by ORCHARD. 01-Sep-2009: Contacted by JYOTI. Arabidopsis cDNA library made from aerial parts of the palnt including flowers. Plant Physiol. (0032-0889) 2004 orchard Only protein-protein interactions imex curation asf1b_arath Histone chaperone ASF1B ASF1B SGA01 SGA1 Anti-silencing function protein 1-like protein b Silencing group A protein 1 F16F17.110 At5g38110 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSSINITNVTVLDNPAPFVNPFQFEISYECLTSLKDDLEWKLIYVGSAEDETYDQVLESVLVGPVNVGNYRFVLQADSPDPLKIREEDIIGVTVLLLTCSYMDQEFIRVGYYVNNDYDDEQLREEPPTKVLIDKVQRNILTDKPRVTKFPINFHPENEQTLGDGPAPTEPFADSVVNGEAPVFLEQPQKLQEIEQFDDSDVNGEAIALLDQPQNLQET 80E29847E65E25FC tki1_arath TSL-kinase interacting protein 1 TKI1 Myb-related protein TKI1 T1J8.14 At2g36960 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MLKQFTHYCEMQAELIPEGPNGEGRLSNQNSNPNLLSSASISITQFPAKKPTRQWAAWTHQEEESFFTALRQVGKNFEKITSRVQSKNKDQVRHYYYRLVRRMNKLLGPDLSLDAKNPKDTNAAMLRWWSLLEKYSCKASKLHLKPRRFKLFIEALEHQLLKDRRKSIRKRTCQGENLSSASLGNISSHSRERGLDNRPFKLILSDGQNVKKLGPGRASTKHGESLSVNLGDEKEDTAFGRGGRQRRKQGYRKWEKAAIDGVSLVADAAEHLERTSIDKDMDDQTDLGPTRYLTGKSPLSLCSAGDVPLSDANMQFSAKLKLQLFPIDECTRRSLEMDKHNPHLELTLSNRKKISSVLEHLNRKWGSSSCATGELLLFPYNARKETVTCHQRWTHDSFLSAAEVHSMVGSPSVFRLRYGWFVHDASGSIISQVPTSDPCPSLEDDMNVDRLNEVNMLLTESGPLSVHSTAEQTTSVEPSQGLVCASGVHDRPARSRDDYEPASTSITPLEHLSGGNAQSPGEWADSLTNISIGDLLSEVPDDIDSDGVDPPATEGSHYLLRDVPFTSDSFDAAIAAHILRHQNKPSAQLPLTSGSSSLWDDEETRDAFSFQKNRFANSTELASVASPKGVGRVNGEPSQLVEASSGDEGSYNPHDDGDPMEEGPADPHTMDSPGKTPCGLADVYWPDSLGPLDLDIRSSKYTDDLILSESLGGLSRLIATSLDAFQNCSLFGFDNKKDKSNMV B952325D7102ECCC tsl_arath Serine/threonine-protein kinase TOUSLED TOUSLED TSL F22D1.100 At5g20930 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSDDMVLHFSSNSSNQSDHSLPDKIAKLEARLTGKTPSSAKPPQQQQQQQQQVSLWSSASAAVKVVTSTPPGLSETSISDSDDENTGDFLIRANTKKRQKVQESNNFSVVDHVEPQEAAYDGRKNDAESKTGLDVSKKKQGRGRASSTGRGRGSKTNNDVTKSQFVVAPVSAASQLDASDQKDFRPDGQLRNGECSLQDEDLKSLRAKIAMLEEELRKSRQDSSEYHHLVRNLENEVKDLKDQEQQGKQKTTKVISDLLISVSKTERQEARTKVRNESLRLGSVGVLRTGTIIAETWEDGQMLKDLNAQLRQLLETKEAIERQRKLLKKRQNGDKNDGTDTESGAQEEDIIPDEVYKSRLTSIKREEEAVLRERERYTLEKGLLMREMKRIRDEDGSRFNHFPVLNSRYALLNLLGKGGFSEVYKAYDLVDHRYVACKLHGLNAQWSEEKKQSYIRHANRECEIHKSLVHHHIVRLWDKFHIDMHTFCTVLEYCSGKDLDAVLKATSNLPEKEARIIIVQIVQGLVYLNKKSQKIIHYDLKPGNVLFDEFGVAKVTDFGLSKIVEDNVGSQGMELTSQGAGTYWYLPPECFELNKTPMISSKVDVWSVGVLFYQMLFGKRPFGHDQSQERILREDTIIKAKKVEFPVTRPAISNEAKDLIRRCLTYNQEDRPDVLTMAQDPYLAYSKK 2B14F0E677D78C9B tsl-tkl1 3 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association The yeast strain YD116 was transformed with the TSL bait vector pCD-NQ-TSL, followed by a cDNA library in a Gal4-activation domain (ACT) plasmid, and clones encoding proteins that interact with TSL were selected for by growth on medium lacking Trp, Leu, and Ura. A total of 1 x 106 transformants were screened and putative positives were retested with the bait vector in a different strain, Y153, for activation of aUASGal4::lacZ reporter gene. A number of clones were eliminated as false positives, and several were able to activate transcription alone (data not shown) and also were eliminated. One clone, however, showed strong reporter gene activation that was completely dependent on the presence of the TSL bait vector (Fig. 5). Furthermore, it was shown that the clone, pC-TKI (TSL-kinase interacting protein), did not show activation when cotransformed with other DBD-fusion protein vectors such as lamin (data not shown). So, it appears that the TKI1 gene encodes a polypeptide that specifically interacts with TSL. tsl-tkl1-1 2 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait range sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q39238:p.Lys438Glu mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain K E Wed May 02 18:31:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys438glu' Tue Jun 30 18:14:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys438Glu' physical association physical association TKI1 interacts with active forms of TS Figure 5 tsl-asfb 2 4 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q39238:p.Lys438Glu mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain K E Wed May 02 18:41:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys438glu' Tue Jun 30 15:54:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys438Glu' physical association physical association TSL interacts with Asf1b/SGA1 and TKI1 in different domains. Figure 7 tsl-tkl1-2 2 6 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain Q39238:p.Lys438Glu mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K E Wed May 02 17:00:06 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys438glu' Tue Jun 30 20:04:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys438Glu' 5 unspecified role unspecified role prey prey c-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 23 26 physical association physical association Figs. 5 and 7 TSL interacts approximately fourfold more strongly with the full-length TKI1 protein. tsl-tsl1 1 6 putative self putative self putative self putative self Q39238:p.Lys438Glu mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain K E Wed May 02 18:58:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys438glu' Tue Jun 30 18:26:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys438Glu' region 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast phosphorylation phosphorylation reaction phosphorylation true Autophosphorylated GST-TSL appears at approximately 106 kD. Figure 8 autophosphorylation tsl-asf1b 1 6 enzyme enzyme neutral component neutral component region 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 Q39238:p.Lys438Glu mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain K E Wed May 02 18:46:30 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys438glu' Tue Jun 30 17:34:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys438Glu' yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 4 enzyme target enzyme target substrate substrate neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast phosphorylation phosphorylation reaction phosphorylation TKI1, Asf1b, and histone H3 are substrates of TSL in vitro. Figure 8 phosphorylation of Q9LS09 tsl-tkl1-3 1 6 enzyme enzyme neutral component neutral component region 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 Q39238:p.Lys438Glu mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain K E Wed May 02 17:22:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys438glu' Tue Jun 30 18:16:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys438Glu' yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 5 enzyme target enzyme target substrate substrate neutral component neutral component c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation TKI1, Asf1b, and histone H3 are substrates of TSL in vitro. Figure 8 phosphorylation of Q8LJT8