UniProt UniProt http://www.uniprot.org STIL binding to Polo-box 3 of PLK4 regulates centriole duplication. STIL binding to Polo-box 3 of PLK4 regulates centriole duplication. eLife 2014 imex curation Arquint C., Gabryjonczyk AM., Imseng S., Böhm R., Sauer E., Hiller S., Nigg EA., Maier T. timm.maier@unibas.ch 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference pull down pull down ms participant Identification by mass spectrometry ms participant Arquint C., Gabryjonczyk AM., Imseng S., Böhm R., Sauer E., Hiller S., Nigg EA., Maier T. A U2OS Flp-In T-REx cell linewas deveoped to allow the inducible expression of an S-peptide-EGFP-tagged PLK4 fragment. timm.maier@unibas.ch Accepted 2018-JUN-07 AT 15:22 BST AT 15:22 BST by LUANA eLife 2014 imex curation STIL binding to Polo-box 3 of PLK4 regulates centriole duplication. STIL binding to Polo-box 3 of PLK4 regulates centriole duplication. eLife 2014 imex curation Arquint C., Gabryjonczyk AM., Imseng S., Böhm R., Sauer E., Hiller S., Nigg EA., Maier T. timm.maier@unibas.ch 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Arquint C., Gabryjonczyk AM., Imseng S., Böhm R., Sauer E., Hiller S., Nigg EA., Maier T. A U2OS Flp-In T-REx cell linewas deveoped to allow the inducible expression of an S-peptide-EGFP-tagged PLK4 fragment. timm.maier@unibas.ch Accepted 2018-JUN-07 AT 15:22 BST AT 15:22 BST by LUANA eLife 2014 imex curation STIL binding to Polo-box 3 of PLK4 regulates centriole duplication. STIL binding to Polo-box 3 of PLK4 regulates centriole duplication. eLife 2014 imex curation Arquint C., Gabryjonczyk AM., Imseng S., Böhm R., Sauer E., Hiller S., Nigg EA., Maier T. timm.maier@unibas.ch 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Arquint C., Gabryjonczyk AM., Imseng S., Böhm R., Sauer E., Hiller S., Nigg EA., Maier T. A U2OS Flp-In T-REx cell linewas deveoped to allow the inducible expression of an S-peptide-EGFP-tagged PLK4 fragment. timm.maier@unibas.ch Accepted 2018-JUN-07 AT 15:22 BST AT 15:22 BST by LUANA eLife 2014 imex curation STIL binding to Polo-box 3 of PLK4 regulates centriole duplication. STIL binding to Polo-box 3 of PLK4 regulates centriole duplication. eLife 2014 imex curation Arquint C., Gabryjonczyk AM., Imseng S., Böhm R., Sauer E., Hiller S., Nigg EA., Maier T. timm.maier@unibas.ch 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Arquint C., Gabryjonczyk AM., Imseng S., Böhm R., Sauer E., Hiller S., Nigg EA., Maier T. A U2OS Flp-In T-REx cell linewas deveoped to allow the inducible expression of an S-peptide-EGFP-tagged PLK4 fragment. timm.maier@unibas.ch Accepted 2018-JUN-07 AT 15:22 BST AT 15:22 BST by LUANA eLife 2014 imex curation STIL binding to Polo-box 3 of PLK4 regulates centriole duplication. STIL binding to Polo-box 3 of PLK4 regulates centriole duplication. eLife 2014 imex curation Arquint C., Gabryjonczyk AM., Imseng S., Böhm R., Sauer E., Hiller S., Nigg EA., Maier T. timm.maier@unibas.ch 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference fluorescence imaging fluorescence microscopy fluorescence imaging immunocytochemistry immunocytochemistry Arquint C., Gabryjonczyk AM., Imseng S., Böhm R., Sauer E., Hiller S., Nigg EA., Maier T. A U2OS Flp-In T-REx cell linewas deveoped to allow the inducible expression of an S-peptide-EGFP-tagged PLK4 fragment. timm.maier@unibas.ch Accepted 2018-JUN-07 AT 15:22 BST AT 15:22 BST by LUANA eLife 2014 imex curation STIL binding to Polo-box 3 of PLK4 regulates centriole duplication. STIL binding to Polo-box 3 of PLK4 regulates centriole duplication. eLife 2014 imex curation Arquint C., Gabryjonczyk AM., Imseng S., Böhm R., Sauer E., Hiller S., Nigg EA., Maier T. timm.maier@unibas.ch 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Arquint C., Gabryjonczyk AM., Imseng S., Böhm R., Sauer E., Hiller S., Nigg EA., Maier T. Origin of PLK4 inferred from sequence length of protein described by Westendorf in PMID: 17681131 timm.maier@unibas.ch Accepted 2018-JUN-07 AT 15:22 BST AT 15:22 BST by LUANA eLife 2014 imex curation stil_human SCL-interrupting locus protein STIL SIL TAL-1-interrupting locus protein protein protein human Homo sapiens Human MEPIYPFARPQMNTRFPSSRMVPFHFPPSKCALWNPTPTGDFIYLHLSYYRNPKLVVTEKTIRLAYRHAKQNKKNSSCFLLGSLTADEDEEGVTLTVDRFDPGREVPECLEITPTASLPGDFLIPCKVHTQELCSREMIVHSVDDFSSALKALQCHICSKDSLDCGKLLSLRVHITSRESLDSVEFDLHWAAVTLANNFKCTPVKPIPIIPTALARNLSSNLNISQVQGTYKYGYLTMDETRKLLLLLESDPKVYSLPLVGIWLSGITHIYSPQVWACCLRYIFNSSVQERVFSESGNFIIVLYSMTHKEPEFYECFPCDGKIPDFRFQLLTSKETLHLFKNVEPPDKNPIRCELSAESQNAETEFFSKASKNFSIKRSSQKLSSGKMPIHDHDSGVEDEDFSPRPIPSPHPVSQKISKIQPSVPELSLVLDGNFIESNPLPTPLEMVNNENPPLINHLEHLKPLQPQLYDEKHSPEVEAGEPSLRGIPNQLNQDKPALLRHCKVRQPPAYKKGNPHTRNSIKPSSHNGPSHDIFEKLQTVSAGNVQNEEYPIRPSTLNSRQSSLAPQSQPHDFVFSPHNSGRPMELQIPTPPLPSYCSTNVCRCCQHHSHIQYSPLNSWQGANTVGSIQDVQSEALQKHSLFHPSGCPALYCNAFCSSSSPIALRPQGDMGSCSPHSNIEPSPVARPPSHMDLCNPQPCTVCMHTPKTESDNGMMGLSPDAYRFLTEQDRQLRLLQAQIQRLLEAQSLMPCSPKTTAVEDTVQAGRQMELVSVEAQSSPGLHMRKGVSIAVSTGASLFWNAAGEDQEPDSQMKQDDTKISSEDMNFSVDINNEVTSLPGSASSLKAVDIPSFEESNIAVEEEFNQPLSVSNSSLVVRKEPDVPVFFPSGQLAESVSMCLQTGPTGGASNNSETSEEPKIEHVMQPLLHQPSDNQKIYQDLLGQVNHLLNSSSKETEQPSTKAVIISHECTRTQNVYHTKKKTHHSRLVDKDCVLNATLKQLRSLGVKIDSPTKVKKNAHNVDHASVLACISPEAVISGLNCMSFANVGMSGLSPNGVDLSMEANAIALKYLNENQLSQLSVTRSNQNNCDPFSLLHINTDRSTVGLSLISPNNMSFATKKYMKRYGLLQSSDNSEDEEEPPDNADSKSEYLLNQNLRSIPEQLGGQKEPSKNDHEIINCSNCESVGTNADTPVLRNITNEVLQTKAKQQLTEKPAFLVKNLKPSPAVNLRTGKAEFTQHPEKENEGDITIFPESLQPSETLKQMNSMNSVGTFLDVKRLRQLPKLF DIP protein Q15468 original sequence version: 93 mint homomint 7F15BEDA8717659C ce192_human Centrosomal protein of 192 kDa CEP192 KIAA1569 PP8407 protein protein human Homo sapiens Human MEDFRGIAEESFPSFLTNSLFGNSGILENVTLSSNLGLPVAVSTLARDRSSTDNRYPDIQASYLVEGRFSVPSGSSPGSQSDAEPRERLQLSFQDDDSISRKKSYVESQRLSNALSKQSALQMETAGPEEEPAGATESLQGQDLFNRASPLEQAQDSPIDFHLQSWMNNKEPKIVVLDAGKHFEDKTLKSDLSHTSLLENEKLILPTSLEDSSDDDIDDEMFYDDHLEAYFEQLAIPGMIYEDLEGPEPPEKGFKLPTNGLRQANENGSLNCKFQSENNSSLISLDSHSSETTHKESEESQVICLPGTSNSIGTGDSRRYTDGMLPFSSGTWGTEKEIENLKGIVPDLNSECASKDVLVKTLRAIDVKLNSDNFHDANANRGGFDLTDPVKQGAECPHQNKTVLHMDGCLDTETPTVSIQENVDVASLKPISDSGINFTDAIWSPTCERRTCECHESIEKNKDKTDLPQSVVYQNEEGRWVTDLAYYTSFNSKQNLNVSLSDEMNEDFRSGSEAFDLIAQDEEEFNKEHQFIQEENIDAHNTSVALGDTSWGATINYSLLRKSRSTSDLDKDDASYLRLSLGEFFAQRSEALGCLGGGNNVKRPSFGYFIRSPEKREPIALIRKSDVSRGNLEKEMAHLNHDLYSGDLNEQSQAQLSEGSITLQVEAVESTSQVDENDVTLTADKGKTEDTFFMSNKPQRYKDKLPDSGDSMLRISTIASAIAEASVNTDPSQLAAMIKALSNKTRDKTFQEDEKQKDYSHVRHFLPNDLEKSNGSNALDMEKYLKKTEVSRYESALENFSRASMSDTWDLSLPKEQTTQDIHPVDLSATSVSVRAPEENTAAIVYVENGESENQESFRTINSSNSVTNRENNSAVVDVKTCSIDNKLQDVGNDEKATSISTPSDSYSSVRNPRITSLCLLKDCEEIRDNRENQRQNECVSEISNSEKHVTFENHRIVSPKNSDLKNTSPEHGGRGSEDEQESFRPSTSPLSHSSPSEISGTSSSGCALESFGSAAQQQQPPCEQELSPLVCSPAGVSRLTYVSEPESSYPTTATDDALEDRKSDITSELSTTIIQGSPAALEERAMEKLREKVPFQNRGKGTLSSIIQNNSDTRKATETTSLSSKPEYVKPDFRWSKDPSSKSGNLLETSEVGWTSNPEELDPIRLALLGKSGLSCQVGSATSHPVSCQEPIDEDQRISPKDKSTAGREFSGQVSHQTTSENQCTPIPSSTVHSSVADMQNMPAAVHALLTQPSLSAAPFAQRYLGTLPSTGSTTLPQCHAGNATVCGFSGGLPYPAVAGEPVQNSVAVGICLGSNIGSGWMGTSSLCNPYSNTLNQNLLSTTKPFPVPSVGTNCGIEPWDSGVTSGLGSVRVPEELKLPHACCVGIASQTLLSVLNPTDRWLQVSIGVLSISVNGEKVDLSTYRCLVFKNKAIIRPHATEEIKVLFIPSSPGVFRCTFSVASWPCSTDAETIVQAEALASTVTLTAIAESPVIEVETEKKDVLDFGDLTYGGWKALPLKLINRTHATVPIRLIINANAVAWRCFTFSKESVRAPVEVAPCADVVTRLAGPSVVNHMMPASYDGQDPEFLMIWVLFHSPKKQISSSDILDSAEEFSAKVDIEVDSPNPTPVLRSVSLRARAGIARIHAPRDLQTMHFLAKVASSRKQHLPLKNAGNIEVYLDIKVPEQGSHFSVDPKNLLLKPGEEHEVIVSFTPKDPEACEERILKIFVQPFGPQYEVVLKGEVISSGSKPLSPGPCLDIPSILSNKQFLAWGGVPLGRTQLQKLALRNNSASTTQHLRLLIRGQDQDCFQLQNTFGSEQRLTSNCEIRIHPKEDIFISVLFAPTRLSCMLARLEIKQLGNRSQPGIKFTIPLSGYGGTSNLILEGVKKLSDSYMVTVNGLVPGKESKIVFSVRNTGSRAAFVKAVGFKDSQKKVLLDPKVLRIFPDKFVLKERTQENVTLIYNPSDRGINNKTATELSTVYLFGGDEISRQQYRRALLHKPEMIKQILPEHSVLQNINFVEAFQDELLVTEVYDLPQRPNDVQLFYGSMCKIILSVIGEFRDCISSREFLQPSSKASLESTSDLGASGKHGGNVSLDVLPVKGPQGSPLLSRAARPPLDQLASEEPWTVLPEHLILVAPSPCDMAKTGRFQIVNNSVRLLRFELCWPAHCLTVTPQHGCVAPESKLQILVSPNSSLSTKQSMFPWSGLIYIHCDDGQKKIVKVQIREDLTQVELLTRLTSKPFGILSPVSEPSVSHLVKPMTKPPSTKVEIRNKSITFPTTEPGETSESCLELENHGTTDVKWHLSSLAPPYVKGVDESGDVFRATYAAFRCSPISGLLESHGIQKVSITFLPRGRGDYAQFWDVECHPLKEPHMKHTLRFQLSGQSIEAENEPENACLSTDSLIKIDHLVKPRRQAVSEASARIPEQLDVTARGVYAPEDVYRFRPTSVGESRTLKVNLRNNSFITHSLKFLSPREPFYVKHSKYSLRAQHYINMPVQFKPKSAGKFEALLVIQTDEGKSIAIRLIGEALGKN DIP protein Q8TEP8 original sequence version: 90 CA0A92F264280C3B ce152_human Centrosomal protein of 152 kDa CEP152 KIAA0912 protein protein human Homo sapiens Human MSLDFGSVALPVQNEDEEYDEEDYEREKELQQLLTDLPHDMLDDDLSSPELQYSDCSEDGTDGQPHHPEQLEMSWNEQMLPKSQSVNGYNEIQSLYAGEKCGNVWEENRSKTEDRHPVYHPEEGGDEGGSGYSPPSKCEQTDLYHLPENFRPYTNGQKQEFNNQATNVIKFSDPQWNHFQGPSCQGLEPYNKVTYKPYQSSAQNNGSPAQEITGSDTFEGLQQQFLGANENSAENMQIIQLQVLNKAKERQLENLIEKLNESERQIRYLNHQLVIIKDEKDGLTLSLRESQKLFQNGKEREIQLEAQIKALETQIQALKVNEEQMIKKSRTTEMALESLKQQLVDLHHSESLQRAREQHESIVMGLTKKYEEQVLSLQKNLDATVTALKEQEDICSRLKDHVKQLERNQEAIKLEKTEIINKLTRSLEESQKQCAHLLQSGSVQEVAQLQFQLQQAQKAHAMSANMNKALQEELTELKDEISLYESAAKLGIHPSDSEGELNIELTESYVDLGIKKVNWKKSKVTSIVQEEDPNEELSKDEFILKLKAEVQRLLGSNSMKRHLVSQLQNDLKDCHKKIEDLHQVKKDEKSIEVETKTDTSEKPKNQLWPESSTSDVVRDDILLLKNEIQVLQQQNQELKETEGKLRNTNQDLCNQMRQMVQDFDHDKQEAVDRCERTYQQHHEAMKTQIRESLLAKHALEKQQLFEAYERTHLQLRSELDKLNKEVTAVQECYLEVCREKDNLELTLRKTTEKEQQTQEKIKEKLIQQLEKEWQSKLDQTIKAMKKKTLDCGSQTDQVTTSDVISKKEMAIMIEEQKCTIQQNLEQEKDIAIKGAMKKLEIELELKHCENITKQVEIAVQNAHQRWLGELPELAEYQALVKAEQKKWEEQHEVSVNKRISFAVSEAKEKWKSELENMRKNILPGKELEEKIHSLQKELELKNEEVPVVIRAELAKARSEWNKEKQEEIHRIQEQNEQDYRQFLDDHRNKINEVLAAAKEDFMKQKTELLLQKETELQTCLDQSRREWTMQEAKRIQLEIYQYEEDILTVLGVLLSDTQKEHISDSEDKQLLEIMSTCSSKWMSVQYFEKLKGCIQKAFQDTLPLLVENADPEWKKRNMAELSKDSASQGTGQGDPGPAAGHHAQPLALQATEAEADKKKVLEIKDLCCGHCFQELEKAKQECQDLKGKLEKCCRHLQHLERKHKAVVEKIGEENNKVVEELIEENNDMKNKLEELQTLCKTPPRSLSAGAIENACLPCSGGALEELRGQYIKAVKKIKCDMLRYIQESKERAAEMVKAEVLRERQETARKMRKYYLICLQQILQDDGKEGAEKKIMNAASKLATMAKLLETPISSKSQSKTTQSALPLTSEMLIAVKKSKRNDVNQKIPCCIESKSNSVNTITRTLCEQAPKRRAACNLQRLLENSEHQSIKHVGSKETHLEFQFGDGSCKHLNSLPRNVSPEFVPCEGEGGFGLHKKKDLLSDNGSESLPHSAAYPFLGTLGNKPSPRCTPGPSESGCMHITFRDSNERLGLKVYKCNPLMESENAASEKSQGLDVQEPPVKDGGDLSDCLGWPSSSATLSFDSREASFVHGRPQGTLEIPSESVKSKQFSPSGYLSDTEESNMICQTMKCQRYQTPYLSEETTYLEPGKISVNCGHPSRHKADRLKSDFKKLSSTLPSSVCQQPSRKLIVPLSSQQDSGFDSPFVNLD DIP protein O94986 original sequence version: 120 DIP protein O94986 original sequence version: 107 1727BDA921E1F6BD plk4_human Serine/threonine-protein kinase PLK4 Polo-like kinase 4 Serine/threonine-protein kinase Sak Serine/threonine-protein kinase 18 PLK4 SAK STK18 protein protein human Homo sapiens Human MATCIGEKIEDFKVGNLLGKGSFAGVYRAESIHTGLEVAIKMIDKKAMYKAGMVQRVQNEVKIHCQLKHPSILELYNYFEDSNYVYLVLEMCHNGEMNRYLKNRVKPFSENEARHFMHQIITGMLYLHSHGILHRDLTLSNLLLTRNMNIKIADFGLATQLKMPHEKHYTLCGTPNYISPEIATRSAHGLESDVWSLGCMFYTLLIGRPPFDTDTVKNTLNKVVLADYEMPSFLSIEAKDLIHQLLRRNPADRLSLSSVLDHPFMSRNSSTKSKDLGTVEDSIDSGHATISTAITASSSTSISGSLFDKRRLLIGQPLPNKMTVFPKNKSSTDFSSSGDGNSFYTQWGNQETSNSGRGRVIQDAEERPHSRYLRRAYSSDRSGTSNSQSQAKTYTMERCHSAEMLSVSKRSGGGENEERYSPTDNNANIFNFFKEKTSSSSGSFERPDNNQALSNHLCPGKTPFPFADPTPQTETVQQWFGNLQINAHLRKTTEYDSISPNRDFQGHPDLQKDTSKNAWTDTKVKKNSDASDNAHSVKQQNTMKYMTALHSKPEIIQQECVFGSDPLSEQSKTRGMEPPWGYQNRTLRSITSPLVAHRLKPIRQKTKKAVVSILDSEEVCVELVKEYASQEYVKEVLQISSDGNTITIYYPNGGRGFPLADRPPSPTDNISRYSFDNLPEKYWRKYQYASRFVQLVRSKSPKITYFTRYAKCILMENSPGADFEVWFYDGVKIHKTEDFIQVIEKTGKSYTLKSESEVNSLKEEIKMYMDHANEGHRICLALESIISEEERKTRSAPFFPIIIGRKPGSTSSPKALSPPPSVDSNYPTRERASFNRMVMHSAASPTQAPILNPSMVTNEGLGLTTTASGTDISSNSLKDCLPKSAQLLKSVFVKNVGWATQLTSGAVWVQFNDGSQLVVQAGVSSISYTSPNGQTTRYGENEKLPDYIKQKLQCLSSILLMFSNPTPNFH DIP protein O00444 original sequence version: 88 DIP protein O00444-1 original sequence version: 93 DIP protein O00444 original sequence version: 145 DIP protein O00444-1 original sequence version: 145 DIP protein O00444 original sequence version: 132 DIP protein O00444-1 original sequence version: 132 DIP protein O00444 original sequence version: 135 4D56F5FD983211A6 plk4-cep152-11 1 7 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 8 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region s tag s tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association nfa plk4-stil-1 5 7 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell 10 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization Fig. 1A As 3D-SIM imaging of U2OS cells revealed extensive co-localization of STIL and PLK4 at the proximal end of daughter centrioles (Figure 1A), we asked whether the two proteins depend on each other for recruitment to this site. plk4-stil-2 3 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined coiled-coil region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell O00444:p.[Ser285Ala;Thr289Ala] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain S A certain certain sequence position certain certain certain sequence position certain T A Wed May 02 17:40:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser285ala,thr289ala' Tue Jun 30 19:08:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser285Ala;Thr289Ala]' region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figs. 1D, 2B, 2C, 2E, Fig1 - figure supplement 2B To confirm the interaction of STIL with PLK4, we transfected HEK293T cells with myc- and FLAG-tagged versions of both proteins and performed co-immunoprecipitation experiments. The ser285ala,thr289ala PLK4 mutant is a non-degradable form of the protein. stil-plk4-1 3 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 1D To confirm the interaction of STIL with PLK4, we transfected HEK293T cells with myc- and FLAG-tagged versions of both proteins and performed co-immunoprecipitation experiments. plk4-stil-3 4 7 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig. 1E Moreover, we detected endogenous STIL along with Cep152 in an immunoprecipitate of myc-PLK4, which had been isolated from a U2OS T-REx cell line (Figure 1E). plk4-stil-4 3 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined coiled-coil region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 2D To confirm the interaction of STIL with PLK4, we transfected HEK293T cells with myc- and FLAG-tagged versions of both proteins and performed co-immunoprecipitation experiments. stil-stil-1 3 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig3 -Supp. 1B To confirm the interaction of STIL with PLK4, we transfected HEK293T cells with myc- and FLAG-tagged versions of both proteins and performed co-immunoprecipitation experiments. stil-stil-2 3 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined coiled-coil region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 3 - Supp. 1D stil-stil-3 3 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined coiled-coil region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 3- Supp 1C To confirm the interaction of STIL with PLK4, we transfected HEK293T cells with myc- and FLAG-tagged versions of both proteins and performed co-immunoprecipitation experiments. plk4-stil-5 3 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Fig. 4B To confirm the interaction of STIL with PLK4, we transfected HEK293T cells with myc- and FLAG-tagged versions of both proteins and performed co-immunoprecipitation experiments. plk4-stil-6 3 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 73 77 76 73 physical association physical association Fig. 4C, 4D To confirm the interaction of STIL with PLK4, we transfected HEK293T cells with myc- and FLAG-tagged versions of both proteins and performed co-immunoprecipitation experiments. plk4-stil-7 6 7 enzyme target enzyme target substrate substrate unspecified role unspecified role purified purified region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. 10 enzyme enzyme unspecified role unspecified role n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst 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 certain certain sequence position certain certain certain sequence position certain O00444:p.Asp154Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain D A Wed May 02 14:56:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp154ala' Tue Jun 30 15:45:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp154Ala' ecoli Escherichia coli (strain K12) physical association physical association Fig1. - figure supplement 2 To confirm the interaction of STIL with PLK4, we transfected HEK293T cells with myc- and FLAG-tagged versions of both proteins and performed co-immunoprecipitation experiments. plk4-stil-8 2 7 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 10 unspecified role unspecified role neutral component neutral component purified purified 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) 87 89 direct interaction direct interaction Fig. 4E To quantify the PLK4-PB3/STIL-CC interaction we determined the binding affinity of PLK4-PB3 and STIL-CC using isothermal titration calorimetry. plk4-stil-9 3 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined coiled-coil region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q15468:p.[Leu733Tyr;Leu743Tyr] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain L Y certain certain sequence position certain certain certain sequence position certain L Y Wed May 02 15:07:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu733tyr,leu743tyr' Tue Jun 30 18:06:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu733Tyr;Leu743Tyr]' Q15468:p.[Leu733Asp;Leu743Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain L D certain certain sequence position certain certain certain sequence position certain L D Wed May 02 16:55:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu733asp,leu743asp' Tue Jun 30 19:30:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu733Asp;Leu743Asp]' Q15468:p.[Gln737Lys;Gln739Glu] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain Q K certain certain sequence position certain certain certain sequence position certain Q E Thu Apr 12 11:13:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln737glu,gln739lys' Wed May 02 18:39:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln737lys,gln739glu' Tue Jun 30 16:37:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Gln737Lys;Gln739Glu]' Q15468:p.[Leu733Asp;Leu736Glu;Leu743Asp] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain L D certain certain sequence position certain certain certain sequence position certain L E certain certain sequence position certain certain certain sequence position certain L D Wed May 02 15:23:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu733asp,leu736glu,leu743asp' Tue Jun 30 17:41:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu733Asp;Leu736Glu;Leu743Asp]' Q15468:p.[Leu733Asp;Ile740Lys;Leu743Asp] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain L D certain certain sequence position certain certain certain sequence position certain I K certain certain sequence position certain certain certain sequence position certain L D Wed May 02 19:16:48 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu733asp,ile740lys,leu743asp' Tue Jun 30 15:53:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu733Asp;Ile740Lys;Leu743Asp]' Q15468:p.[Leu733Asp;Leu736Glu;Ile740Lys;Leu743Asp] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain L D certain certain sequence position certain certain certain sequence position certain L E certain certain sequence position certain certain certain sequence position certain I K certain certain sequence position certain certain certain sequence position certain L D Wed May 02 15:00:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu733asp,leu736glu,ile740lys,leu743asp' Tue Jun 30 19:21:57 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu733Asp;Leu736Glu;Ile740Lys;Leu743Asp]' Q15468:p.[Leu733Glu;Ile740Glu;Leu743Glu] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain L E certain certain sequence position certain certain certain sequence position certain I E certain certain sequence position certain certain certain sequence position certain L E Wed May 02 17:50:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu733glu,ile740glu,leu743glu' Tue Jun 30 16:12:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu733Glu;Ile740Glu;Leu743Glu]' region s tag s tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Fig. 6D To confirm the interaction of STIL with PLK4, we transfected HEK293T cells with myc- and FLAG-tagged versions of both proteins and performed co-immunoprecipitation experiments.