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 C. elegans Tousled-like kinase contributes to chromosome segregation as a substrate and regulator of the Aurora B kinase. The C. elegans Tousled-like kinase contributes to chromosome segregation as a substrate and regulator of the Aurora B kinase. Curr. Biol. (0960-9822) 2005 imex curation Han Z., Riefler GM., Saam JR., Mango SE., Schumacher JM. jschumac@mdanderson.org 26-Sep-2011: Contacted by JYOTI. a C. elegans cDNA library - a gift from R. Barstead. jyoti 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 Han Z., Riefler GM., Saam JR., Mango SE., Schumacher JM. jschumac@mdanderson.org Accepted 2011-JUL-01 by JYOTI 26-Sep-2011: Contacted by JYOTI. a C. elegans cDNA library - a gift from R. Barstead. Curr. Biol. (0960-9822) 2005 jyoti Only protein-protein interactions imex curation The C. elegans Tousled-like kinase contributes to chromosome segregation as a substrate and regulator of the Aurora B kinase. The C. elegans Tousled-like kinase contributes to chromosome segregation as a substrate and regulator of the Aurora B kinase. Curr. Biol. (0960-9822) 2005 imex curation Han Z., Riefler GM., Saam JR., Mango SE., Schumacher JM. jschumac@mdanderson.org 26-Sep-2011: Contacted by JYOTI. a C. elegans cDNA library - a gift from R. Barstead. jyoti Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Han Z., Riefler GM., Saam JR., Mango SE., Schumacher JM. Interactions involving MYBP (myelin base protein) cannot be included as no species is given for this protein. (no C. elegans protein exists). Interactions involving (human) Histone H3 have not been included as the identity of the protein is not stated. jschumac@mdanderson.org Accepted 2011-JUL-01 by JYOTI 26-Sep-2011: Contacted by JYOTI. a C. elegans cDNA library - a gift from R. Barstead. Curr. Biol. (0960-9822) 2005 jyoti Only protein-protein interactions imex curation The C. elegans Tousled-like kinase contributes to chromosome segregation as a substrate and regulator of the Aurora B kinase. The C. elegans Tousled-like kinase contributes to chromosome segregation as a substrate and regulator of the Aurora B kinase. Curr. Biol. (0960-9822) 2005 imex curation Han Z., Riefler GM., Saam JR., Mango SE., Schumacher JM. jschumac@mdanderson.org 26-Sep-2011: Contacted by JYOTI. a C. elegans cDNA library - a gift from R. Barstead. jyoti Only protein-protein interactions caeel-embryo Caenorhabditis elegans embryo embryo Embryo Defined as the development stage prior to the initiation of tissue differentiation. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Han Z., Riefler GM., Saam JR., Mango SE., Schumacher JM. jschumac@mdanderson.org Accepted 2011-JUL-01 by JYOTI 26-Sep-2011: Contacted by JYOTI. a C. elegans cDNA library - a gift from R. Barstead. Curr. Biol. (0960-9822) 2005 jyoti Only protein-protein interactions imex curation air2_caeel Aurora/IPL1-related protein kinase 2 stu-7 B0207.4 Serine/threonine-protein kinase aurora-B air-2 protein protein caeel Caenorhabditis elegans MENKPPVINLPEKETVNTPQKGGKFTINDFEIGRPLGKGKFGSVYLARTKTGHFHVAIKVLFKSQLISGGVEHQLEREIEIQSHLNHPNIIKLYTYFWDAKKIYLVLEYAPGGEMYKQLTVSKRFSEPTAAKYMYEIADALSYCHRKNVIHRDIKPENLLIGSQGELKIGDFGWSVHAPSNKRQTMCGTMDYLPPEMVNGADHSDAVDLWAIGVLCYEFLVGKPPFEHEDQSKTYAAIKAARFTYPDSVKKGARDLIGRLLVVDPKARCTLEQVKEHYWIQGMMEAKIRAEKQQKIEKEASLRNH 1635EB60D2E14011 asf1_caeel Probable histone chaperone asf-1 unc-85 Anti-silencing function protein 1 asf-1 F10G7.3 protein protein caeel Caenorhabditis elegans MASRVNIVQVQILDNPAMFVDKFKLEITFEVFEHLPHDLEWELVYVGSGTSRDFDQVLDSALVGPIPEGRHKFVFDADHPDISKIPVDDIVGVSVLLLRCKYNDQEFINMGWFVANEYTEEELKENPPSQPLIEKLSRKVETEDLRITTFPIRWTDEDPVAEPVEDEANRVFAEDDLMPLNDDGQEDDDEEEEDDDEMEANAEEVDLNESFNERLANALDGAEQKGADEKMEDDGANEDVDMADDEPGVQINTDTKVPESMAEPLSDKTNNEMVQ tlk1_caeel Serine/threonine-protein kinase tousled-like 1 tlk-1 Tousled-like kinase 1 C07A9.3 protein protein caeel Caenorhabditis elegans MSMLSMDGIVAGGGSSSGGGERSFVLEQKMFNTGPQNKALPTVQSSGSSSNHAPIVGESPLGTVSSTMATGDTGRVGNVTYMSSGMLGATQFMPQNSSHPSTSVMMQQVPPQNGGATRSSPTEMQQCMQAMSEDSIEMRDYNSGVHHMHPHQMQMQQQQQHHQQQYNMSYHNHQQQMQQMHYHQQQQQYQQQQAQHHQMYAPQIQQQQQQPQQQSQQQSAQQPQQSSAALQHVNESSNLSSAGSISDREPEQHGGTPQRPTAPQSSTATDKKTRKRRKAGPTEDQATPKQERKITEFMKVGGEVASGNSVARCLLTEYHQNQGSPKRQPAVQQNGSNSYDSQQQQPQMNQHEMQNSYWGVATPSLGVNNRGTPTPTQQQHYSSDSNSNSNQSPPGQGNQSGRMVRTIDEETQTDSSLSQANPQNADEVAKMNRIIEDHRRQIEELNSKNSLERRKNEASKETIKRLLIDKNQIERKALRDKTAADSPRIGCFKTTRTGDSFRDQWVDGWAFAEMDKKTEQINAERNEIASASALLKKRKPLGIGKEPKRPQAVNSQNDSNGMQPSTSSNTNGDDAIFRRPEEPKEIQYQEYIELDEIYKLRREHLRKEETDLSMEKERLEKEKQHHVRELKRASNESASQFNDHRLLHKRYLMLNLLGKGGFSEVWKAFDIEENRYVACKIHHVNKDWKEEKKANYVKHAMREKDIHKSLDHCRIVKQYDLLTIDNHSFCTVLEYVPGNDLDFYLKQNRSISEKEARSIIMQVVSALVYLNEKSTPIIHYDLKPANILLESGNTSGAIKITDFGLSKIMEGESDDHDLGIELTSQFAGTYWYLPPETFIVPPPKITCKVDVWSIGVIFYQCIYGKKPFGNDLTQQKILEYNTIINAREVSFPSKPQVSSAAQDFIRRCLQYRKEDRADVFELAKHELFRPRGAIRASVAGSLLLVIITLCQHSNRSEDRLCVSNV g5ee37_caeel icp-1 Y39G10AR.13 CELE_Y39G10AR.13 protein protein caeel Caenorhabditis elegans MPPKKRAVSRKPAEPTIKLFSSIPKISSIFDEVSEAHDDAAAKVFADLYKRMSSITEELATPQFSAEQLLNAVLKPGRIDAEFVELISRQKAKPAQPSSSSHRGDDDMETDEVETDQAGPITPRAGSVATDGDFEEDILPVTLKRMQQMSISKKKTEGSISRNAAYSGTPRRNPPREAHNQATPRNIFSSTPGRMAGTPGRAAVVPRTPHRVIPFNDAEKDPNARMKHADELRMKELEKKREKARKDEEKRNAVMERRKEQERVRQEKLDQLKQKEERNANLAKLHNSAKSPTRARLMQDHAPNKAASRKIFPTAESTSTPGRGPAKKGKVEILIGSDGQKTAPVAQPTVELSPSRELQRNGSRQVKMEPMSCDDSTPPARQHKPKAKATKRSAHAASSSTSNVEAAAALAALQEQQRIQLEQEQYLLQLAEDERRKREQKEMKEKAEKERQLRAREEQEHFLAVQREEQAQLEKQREREEAELQATLARCAEKQARQEALRKTPPPAAYEMTPPRTYQNNSKNDYGLNDLNSDDETDQEDDPRKDVPAWAEFAVVRENVRRHAINPPFDVAQFFGSIGKPNLKEIFSDAVKVKKRGSSAVWKKSPSTLNTSSRTILQDISE 7E055664F8D38DEB air_2-tlk1-1 3 4 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association Fig 1 air_2-air_2-1 2 4 self self self self 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 ecoli Escherichia coli (strain K12) 4 self self self self 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 ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Fig 2A autophosphorylation A marked increase in AIR-2 autophosphorylation and ICP-1 phosphorylation was seen in kinase reactions with all three fusion proteins (AIR-2, ICP-1, and TLK-1KD). air_2-icp1 2 4 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 ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate 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 ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Fig 2A, lane 2 autophosphorylation of AIR-2 and phosphorylation of ICP-1. A marked increase in AIR-2 autophosphorylation and ICP-1 phosphorylation was seen in kinase reactions with all three fusion proteins (AIR-2, ICP-1, and TLK-1KD). air_2-icp_1-3 2 6 enzyme target enzyme target substrate substrate neutral component neutral component P34314:p.Ser634Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain S A Wed May 02 14:55:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser634ala' Tue Jun 30 18:22:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser634Ala' region mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P34314:p.Asp802Ala mutation mutation certain certain sequence position certain certain certain sequence position certain D A Wed May 02 19:42:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp802ala' Tue Jun 30 16:14:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp802Ala' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate ancillary ancillary 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 ecoli Escherichia coli (strain K12) 4 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 ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Fig 2A, lane 4. Fig 2D/E. autophosphorylation of AIR-2 and phosphorylation of ICP-1 and TLK-1 KD. "TLK-1 KD" has been entered as a D802A mutation, but this is not explicitly stated in the text. A marked increase in AIR-2 autophosphorylation and ICP-1 phosphorylation was seen in kinase reactions with all three fusion proteins (AIR-2, ICP-1, and TLK-1KD). Mutation of TLK-1 Serine 634 to alanine (S634A) abolished AIR-2-mediated phosphorylation of TLK-1 fragment 454–650 (Figure 2E, lanes 2 and 3). However, the S634A mutation did not completely inhibit AIR-2 phosphorylation when incorporated into E. coli fusion proteins corresponding to full-length TLK-1. asf1-tlk1 2 5 enzyme target enzyme target substrate substrate 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 6 enzyme enzyme neutral component neutral component P34314:p.Ser634Glu mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain S E Wed May 02 15:56:21 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser634glu' Tue Jun 30 15:50:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser634Glu' P34314:p.Ser634Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain S A Wed May 02 15:35:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser634ala' Tue Jun 30 18:08:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser634Ala' mbp_or_ gst-tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Fig 4 Auto phosphorylation of TLK-1 and phosphorylation of ASF1 and MYBP. TLK-1(S634E) protein showed consistently higher levels of kinase activity than TLK-1 or TLK-1(S634A). ASF-1 phosphorylation was increased when AIR-2/ICP-1 was added to TLK-1. air_2-tlk1-3 1 6 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association See Supplement, paragraph 1. tlk1-tlk1-2 2 6 self self self self mbp_or_ gst-tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) palmitoylation palmitoylation reaction palmitoylation true Fig S1 Auto phosphorylation of TLK-1.