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/ http://www.ebi.ac.uk/ han-2005-2 The C. elegans Tousled-like kinase contributes to chromosome segregation as a substrate and regulator of the Aurora B kinase. 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 a C. elegans cDNA library - a gift from R. Barstead. Curr. Biol. (0960-9822) 2005 Only protein-protein interactions imex curation han-2005-1 The C. elegans Tousled-like kinase contributes to chromosome segregation as a substrate and regulator of the Aurora B kinase. 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 a C. elegans cDNA library - a gift from R. Barstead. Curr. Biol. (0960-9822) 2005 Only protein-protein interactions imex curation han-2005-3 The C. elegans Tousled-like kinase contributes to chromosome segregation as a substrate and regulator of the Aurora B kinase. 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 a C. elegans cDNA library - a gift from R. Barstead. Curr. Biol. (0960-9822) 2005 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 g5ee37_caeel icp-1 Y39G10AR.13 CELE_Y39G10AR.13 protein protein caeel Caenorhabditis elegans MPPKKRAVSRKPAEPTIKLFSSIPKISSIFDEVSEAHDDAAAKVFADLYKRMSSITEELATPQFSAEQLLNAVLKPGRIDAEFVELISRQKAKPAQPSSSSHRGDDDMETDEVETDQAGPITPRAGSVATDGDFEEDILPVTLKRMQQMSISKKKTEGSISRNAAYSGTPRRNPPREAHNQATPRNIFSSTPGRMAGTPGRAAVVPRTPHRVIPFNDAEKDPNARMKHADELRMKELEKKREKARKDEEKRNAVMERRKEQERVRQEKLDQLKQKEERNANLAKLHNSAKSPTRARLMQDHAPNKAASRKIFPTAESTSTPGRGPAKKGKVEILIGSDGQKTAPVAQPTVELSPSRELQRNGSRQVKMEPMSCDDSTPPARQHKPKAKATKRSAHAASSSTSNVEAAAALAALQEQQRIQLEQEQYLLQLAEDERRKREQKEMKEKAEKERQLRAREEQEHFLAVQREEQAQLEKQREREEAELQATLARCAEKQARQEALRKTPPPAAYEMTPPRTYQNNSKNDYGLNDLNSDDETDQEDDPRKDVPAWAEFAVVRENVRRHAINPPFDVAQFFGSIGKPNLKEIFSDAVKVKKRGSSAVWKKSPSTLNTSSRTILQDISE tlk1_caeel Serine/threonine-protein kinase tousled-like 1 tlk-1 Tousled-like kinase 1 C07A9.3 protein protein caeel Caenorhabditis elegans MSMLSMDGIVAGGGSSSGGGERSFVLEQKMFNTGPQNKALPTVQSSGSSSNHAPIVGESPLGTVSSTMATGDTGRVGNVTYMSSGMLGATQFMPQNSSHPSTSVMMQQVPPQNGGATRSSPTEMQQCMQAMSEDSIEMRDYNSGVHHMHPHQMQMQQQQQHHQQQYNMSYHNHQQQMQQMHYHQQQQQYQQQQAQHHQMYAPQIQQQQQQPQQQSQQQSAQQPQQSSAALQHVNESSNLSSAGSISDREPEQHGGTPQRPTAPQSSTATDKKTRKRRKAGPTEDQATPKQERKITEFMKVGGEVASGNSVARCLLTEYHQNQGSPKRQPAVQQNGSNSYDSQQQQPQMNQHEMQNSYWGVATPSLGVNNRGTPTPTQQQHYSSDSNSNSNQSPPGQGNQSGRMVRTIDEETQTDSSLSQANPQNADEVAKMNRIIEDHRRQIEELNSKNSLERRKNEASKETIKRLLIDKNQIERKALRDKTAADSPRIGCFKTTRTGDSFRDQWVDGWAFAEMDKKTEQINAERNEIASASALLKKRKPLGIGKEPKRPQAVNSQNDSNGMQPSTSSNTNGDDAIFRRPEEPKEIQYQEYIELDEIYKLRREHLRKEETDLSMEKERLEKEKQHHVRELKRASNESASQFNDHRLLHKRYLMLNLLGKGGFSEVWKAFDIEENRYVACKIHHVNKDWKEEKKANYVKHAMREKDIHKSLDHCRIVKQYDLLTIDNHSFCTVLEYVPGNDLDFYLKQNRSISEKEARSIIMQVVSALVYLNEKSTPIIHYDLKPANILLESGNTSGAIKITDFGLSKIMEGESDDHDLGIELTSQFAGTYWYLPPETFIVPPPKITCKVDVWSIGVIFYQCIYGKKPFGNDLTQQKILEYNTIINAREVSFPSKPQVSSAAQDFIRRCLQYRKEDRADVFELAKHELFRPRGAIRASVAGSLLLVIITLCQHSNRSEDRLCVSNV asf1_caeel Probable histone chaperone asf-1 unc-85 Anti-silencing function protein 1 asf-1 F10G7.3 protein protein caeel Caenorhabditis elegans MASRVNIVQVQILDNPAMFVDKFKLEITFEVFEHLPHDLEWELVYVGSGTSRDFDQVLDSALVGPIPEGRHKFVFDADHPDISKIPVDDIVGVSVLLLRCKYNDQEFINMGWFVANEYTEEELKENPPSQPLIEKLSRKVETEDLRITTFPIRWTDEDPVAEPVEDEANRVFAEDDLMPLNDDGQEDDDEEEEDDDEMEANAEEVDLNESFNERLANALDGAEQKGADEKMEDDGANEDVDMADDEPGVQINTDTKVPESMAEPLSDKTNNEMVQ air_2-air_2-1 1136375 n/a 1136378 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 false ecoli Escherichia coli (strain K12) n/a 1136378 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 false ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false 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 1136375 n/a 1136378 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 false ecoli Escherichia coli (strain K12) n/a 1136379 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 false ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false 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 1136375 n/a 1136380 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 false 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 false P34314:p.Asp802Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1136379 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 false ecoli Escherichia coli (strain K12) n/a 1136378 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 false ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false 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 1136375 n/a 1136381 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 false n/a 1136380 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 false P34314:p.Ser634Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false mbp_or_ gst-tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false 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. tlk1-tlk1-2 1136375 n/a 1136380 self self self self mbp_or_ gst-tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) palmitoylation palmitoylation reaction palmitoylation false true false Fig S1 Auto phosphorylation of TLK-1. air_2-tlk1-1 1136376 n/a 1136378 unspecified role unspecified role bait bait n/a 1136380 unspecified role unspecified role prey prey physical association physical association false false false Fig 1 air_2-tlk1-3 1136377 n/a 1136380 unspecified role unspecified role prey prey n/a 1136378 unspecified role unspecified role bait bait physical association physical association false false false See Supplement, paragraph 1.