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/ LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. Neuron (0896-6273) 2000 imex curation Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu A mouse E10.5 total embryonic cDNA library was used. orchard Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu Accepted 2009-SEP-27 by ORCHARD. A mouse E10.5 total embryonic cDNA library was used. Neuron (0896-6273) 2000 orchard Only protein-protein interactions imex curation LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. Neuron (0896-6273) 2000 imex curation Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu A mouse E10.5 total embryonic cDNA library was used. 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 Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu Accepted 2009-SEP-27 by ORCHARD. A mouse E10.5 total embryonic cDNA library was used. Neuron (0896-6273) 2000 orchard Only protein-protein interactions imex curation LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. Neuron (0896-6273) 2000 imex curation Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu A mouse E10.5 total embryonic cDNA library was used. orchard Only protein-protein interactions 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 Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu Accepted 2009-SEP-27 by ORCHARD. A mouse E10.5 total embryonic cDNA library was used. Neuron (0896-6273) 2000 orchard Only protein-protein interactions imex curation LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. Neuron (0896-6273) 2000 imex curation Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu A mouse E10.5 total embryonic cDNA library was used. orchard Only protein-protein interactions 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 bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu Accepted 2009-SEP-27 by ORCHARD. A mouse E10.5 total embryonic cDNA library was used. Neuron (0896-6273) 2000 orchard Only protein-protein interactions imex curation LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. Neuron (0896-6273) 2000 imex curation Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu A mouse E10.5 total embryonic cDNA library was used. orchard Only protein-protein interactions chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu Accepted 2009-SEP-27 by ORCHARD. A mouse E10.5 total embryonic cDNA library was used. Neuron (0896-6273) 2000 orchard Only protein-protein interactions imex curation LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. Neuron (0896-6273) 2000 imex curation Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu A mouse E10.5 total embryonic cDNA library was used. 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 Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu Accepted 2009-SEP-27 by ORCHARD. A mouse E10.5 total embryonic cDNA library was used. Neuron (0896-6273) 2000 orchard Only protein-protein interactions imex curation LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. Neuron (0896-6273) 2000 imex curation Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu A mouse E10.5 total embryonic cDNA library was used. orchard Only protein-protein interactions 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 western blot western blot Immuno blot Immuno blot Feng Y., Olson EC., Stukenberg PT., Flanagan LA., Kirschner MW., Walsh CA. Apoptosis - Interactions involving proteins with a function related to apoptosis cwalsh@caregroup.harvard.edu Accepted 2009-SEP-27 by ORCHARD. A mouse E10.5 total embryonic cDNA library was used. Neuron (0896-6273) 2000 orchard Only protein-protein interactions imex curation dyl1_mouse Dynein light chain 1, cytoplasmic Dynll1 Dlc1 Dnclc1 Dncl1 Dynein light chain LC8-type 1 8 kDa dynein light chain Protein inhibitor of neuronal nitric oxide synthase protein protein mouse Mus musculus Mouse MCDRKAVIKNADMSEEMQQDSVECATQALEKYNIEKDIAAHIKKEFDKKYNPTWHCIVGRNFGSYVTHETKHFIYFYLGQVAILLFKSG F5E7647D092BEB3A cp110_mouse Centriolar coiled-coil protein of 110 kDa Cep110 Ccp110 Kiaa0419 Cp110 Centrosomal protein of 110 kDa protein protein mouse Mus musculus Mouse MEEYEEFCEKALGRAQEASLSTGSFLPAQAESVSLIRFHGVAVLSPLLTIEKRKKIQEEKQKALDVQSRKQANRKKALLTRVQEILENVQVRKAPNASDFDQWATETIYSNPEVTDLNVPVRVPNSLPSPTEHCTSVKLEKITGLLPVNNEDQQTPKRVGLPGDSEVSGSLRQCESPESRQAEDGAALRLSSASPQETIISDVLGKEEQDPSCLAEVTPDPYIMSLQNLMKRSKEYVERELSSRSLRNSLKRSVNETHSDRENDAAKASDCVKEKAPPMPIGRHCGSAIPDKPSLNKSNVLLQGASQASSMGTAGLASFSKIDLPAGAAPPAAPDAGSDFTVIPTFVTENKVKSLKGPYAKLPSPEPSMSPTMHRRHSRSASACQILINNPVNACELSPKGKEEAVDRTAPAAAETTNESETVPKSPTDLTGVCSSNVSATKITSESTREMVVGKPSQRQQALGAHLGNNVTVERSAMEGPFIADDRGAQKVDGTCMAVPKLHELQPSSQCVSSQTLEDVCELKSASLLAKNSCNLQMELNKSYDVKHPSPLLTQTQTSRQQMDTPPVFRGNEQFVDNSFEKVKRRLDLDVDSLQKENCPYIITAGVAEQERDRLLERRYPKGFVHINKNKMLETSPKEGQELLKSKMLAFEEMRKRLEEQHAQQLSLLIAEQEREQEQLQKEIEEQEKMLKEKAVTTDVSDLNSALEWRQRTDSALLETMLSQVDSLQTSNNSGFITSALQYSFGSAGEAPFYLWGSLTSGVTRVSGTRPCGRAQAKWSQVFNPEIHAKFNKITAVAKGFLTRKLMQTDKLKQLRQTVKDTMEFIRSFQSEAPLKRGVVSAQDASLQERVLAQLRAALYGIHDIFFVMDAAERMSILHHDREARKEKLLRQMDKMKSPRVALSVATQKSLDRKKFMKVAEMGMPNKKFLLKQNPSETRVLQPNQGQNAPVHRLLSRQGTPKTSVKGVVQNRQKPSQSRVPNRAPVSGAYAGKTQRKRPNVATI C8D81F5F2FE213A8 q155p7_mouse Cenpf Lek1 protein protein mouse Mus musculus Mouse MSWALEEWKEGLPSRALQKIQELEGQLEKLKKEKQQRQFQLDSLEAALQKQKQKVEDGKTEGADLKRENQRLMEICEHLEKSRQKLSHELQVKESQVNLQESQLSSCKKQIEKLEQELKRCKSEFERSQQVAQSADVSLNPCSTPQKLFATPLTPSSTYEDLKEKYNKEVEERKRLEEEVKALHAKKVSLPVSQATMNHRDIARHQASSSVFPWQQENTPSRLSSDALKTPLRRDGSAAHFLGEEVSPNKSSMKTGRGDCSSLPGEPHSAQLLHQAKAQNQDLKSKMTELELRLQGQEKEMRSQVNKCQDLQLQLEKTKVELIEKERILNKTRDEVVRSTAQYDQAAAKCTTLEQKLKTLTEELSCHRQNAESAKRSLEQRIKEKEKELQEELSRQHQSFQALDSEYTQMKTRLTQELQQVKHLHSTLQLELEKVTSVKQQLERNLEEIRLKLSRAEQALQASQVAENELRRSSEEMKKENSLIRSQSEQRTREVCHLEEELGKVKVSLSKSQNFAEEMKAKNTSQEIMLRDLQEKLNQQENSLTLEKLKLALADLERQRNCSQDLLKKREHHIDQLNNKLNKIEKEFETLLSALELKKKECEELKEEKNQISFWKIDSEKLINQIESEKEILLGKINHLETSLKTQQVSPDSNERIRTLEMERENFTVEIKNLQSMLDSKMVEIKTQKQAYLELQQKSESSDQKHQKEIENMCLKANKLTGQVESLECKLQLLSSEVVTKDQQYQDLRMEYETLRDLLKSRGSSLVTNEDNQRSSEDNQRSSEDNQRGSLAFEQQPAVSDSFANVMGRKGSINSERSDCSVDGGRSPEHIAILQNRVTSLESSLESQNQMNSDLQMRCEELLQIKGEVEENLSKAEQIHQNFVAETNQCISKLQEDAAVHQNIVAETLATLESKEKELQLLKEKLEAQQTEVQKLNKNNCLLEGTLKELQLLSDTLSSEKKEMNSIISLSKKNIEELTQANEALKEVNEALEQEKMNLLQKHEKITSCIAEQERSIAELSDQYKQERLQLLQRCEETEAVLEDLRGNYKTAQENNAKLECMLSECTALCENRKNELEQLKETFAKEQQEFLTKLAFAEEQNRKLMLELEIEQQTVRSEITNTNKHSMSATDGLRQECLTLNEEQNEQQNEVSNLTHENEQLMELTQTKHDSYLAVEPVENSVKATEDEIGKSSSQYQMDIDTKDISLDSYKAQLVHLEALVRILEVQLDQSEEENKKLHLELQTIREELETKSSQDPQSQARTGLKDCDTAEEKYVSMLQELSASQNENAHLQCSLQTAVNKLNELGKMCDVLRVEKLQLESELNDSRTECITATSQMTAEVEKLVSEMKMLNHESALSQNELMKDTSGGEFHDKANHSSVFLTPLDSSNFCEQMTLSSKEVRVHFAELQEKFSCLQSEHKILHDQHCEVSSKMSALRSYVDTLKAENSALSMSLRTLQGDLVKEGEPAAEGGHGLPLSFCGADSPSLTNFGETSFYKDVLEQTGDTCHLSLEGNASANSCDLDEEFSSSLEEETLTEKESPPAPGRTVEGLEVLCQVYLQSLKNLEEKTESQRIMKNKEIEELEQLLSSERKELSCLRKQYLSEKEQWQQKLTSVTLEMESKLAEEKQQTKTLSLELEVARLQLQELDLSSRSLLGTDLESVVRCQNDNYDIKESEVYISETTEKTPKQDTDQTCDKDIQQDLGLETSVTESETTRLTGEGCEEQPPKTNCEAPAEDKTQDCSECISELCSSSNVLVPMDVLEDQGSIQNLQLQKDTLNENLRLLPEVEDWDKKVESLLNEIMEADSKLSLQEVQLKMKIATCIQLEKIVKDLRKEKADLSEKLESLPCNQEVCLRVERSEEDLGFNLDMGANELLSKSTKDNATNTEDNYKEKFLDMERELTRIKSEKANIEHHILSVETNLEVVQAEKLCLERDTESKQKVIIDLKEELFTVISERNRLREELDNVSKESKALDQMSKKMKEKIEELESHQRESLRHIGAVESEVKDKADLIQTLSFNVGELTKDKAHLQEQLQNLQNDSQELSLAIGELEIQIGQLNKEKESLVKESQNFQIKLTESECEKQTISKALEVALKEKGEFAVQLSSAQEEVHQLRRGIEKLSVRIEADEKKHLSAVAKLKESQRESDSLKDTVETLERELERSEENQELAILDSENLKAEVETLKAQKDEMTKSLRIFELDLVTVRTERENLAKQLQEKQSRVSELDERCSSLRRLLEEKEQARVQMEEDSKSAMLMLQMQLKELREEVAALCNDQETLKAQEQSLDQPGEEVHHLKSSIRKLKVHIDADEKKHQNILEQLKESKHHADLLKDRVENLEQELILSEKNMIFQAEKSKAEIQTLKSEIQRMAQNLQDLQLELISTRSENENLIKELKKEQERVSDLETINPSIENLLKDKEQEKVQMKEEAKITVEMLQTQLKELNETVVSLCNDQEVSKTKEQNLGSQVQTLELEKAQLLQDLGEAKNKYIIFQSSVNALTQEVEAGKQKLEKGEKEIRTLKEQLKSQEQLVCKLAQVEGEQELWQKQKLELRNVTMALEQKVQVLQSENNTLQSTYEALQNSHKSLESELGLIKLEKVALVERVSTISGKEAELQRELRDMLQKTTQLSEDYNKEKNRLTEEVEVLREELQNTKAAHLKSVNQLEKELQRAQGKIKLMLKSCRQLEGEKEMLQKELSQLEAAQQQRAGSLVDSNVDEVMTENKALKETLEEKVKEADKYLDKYCSLLISHEELEKAKEILEIEVARLKSRQSRQDLQSSPLLNSSIPGPSPNTSVSEMKSASGQNKASGKRQRSSGVWEHGKRAAPSTAETFSKKSRKSDSKSTRPAEHEQETEFEPEGLPEVVKKGFADIPTGKTSPYILRRTTMATRTSPRFATQKLVGSSPSLGKENVVESSKPTAGGSRSQKVKVVQESSADSHTAFQELPAKSLTASNIPGRNSTESPREGLRAKRAYPASSPAAGPDPTNNENCRVQ 3CB74C4F24537155 nde1_mouse Nuclear distribution protein nudE homolog 1 Nde1 Nude protein protein mouse Mus musculus Mouse MEDSGKTFESEEEETNYWRDLAMTYKQRAENTQEELREFQEGSREYEAELEAQLQQIETRNRDLLSENNRLRMELESVKEKFEMQHSEGYRQISALEDDLAQTKAIKDQLQKYIRELEQANDDLERAKRATIMSLEDFEQRLNQAIERNAFLESELDEKENLLESVQRLKDEARDLRQELAVQQKQDKPRTPMPGSGQAKRTDMAVQATGSVPSTPVAHRGPSSGLNTPGMFRRGLDSSTSGTPLTPAARISALNIVGDLLRKVGALESKLASCRNFMYDQSPSRTSGPASGRGTKNRDGVDRRPGSTSVGDKGSGKRLEFGKPASEPASPALPSAQGVVKLLL C0A339C4F51EF8D9 tbg1_human Tubulin gamma-1 chain TUBG1 TUBG Gamma-1-tubulin Gamma-tubulin complex component 1 protein protein human Homo sapiens Human MPREIITLQLGQCGNQIGFEFWKQLCAEHGISPEGIVEEFATEGTDRKDVFFYQADDEHYIPRAVLLDLEPRVIHSILNSPYAKLYNPENIYLSEHGGGAGNNWASGFSQGEKIHEDIFDIIDREADGSDSLEGFVLCHSIAGGTGSGLGSYLLERLNDRYPKKLVQTYSVFPNQDEMSDVVVQPYNSLLTLKRLTQNADCVVVLDNTALNRIATDRLHIQNPSFSQINQLVSTIMSASTTTLRYPGYMNNDLIGLIASLIPTPRLHFLMTGYTPLTTDQSVASVRKTTVLDVMRRLLQPKNVMVSTGRDRQTNHCYIAILNIIQGEVDPTQVHKSLQRIRERKLANFIPWGPASIQVALSRKSPYLPSAHRVSGLMMANHTSISSLFERTCRQYDKLRKREAFLEQFRKEDMFKDNFDEMDTSREIVQQLIDEYHAATRPDYISWGTQEQ DIP protein P23258 original sequence version: 88 DIP protein P23258 original sequence version: 123 DIP protein P23258 original sequence version: 165 DIP protein P23258 original sequence version: 168 E2A4C0179ED0CFE8 pcnt_mouse Pericentrin Pcnt Pcnt2 protein protein mouse Mus musculus Mouse MEDEQEQRRRKVEAGRAKLANFRQRKTKGDCPNSKKKTAKRKGSAVHASVQEEGSVATPNSELPQGGAVFESPSCSNTLEGTRGASAAQEQEDCELDVTDLQGQQQTQPPPPQTAHSLELEALRLSLNNMHTAQLELTQANLQKEKETALTELREMLNGRRAQELALLQSRQQCELELLREQHAREKEEMALRSGQEAAELKEKLRSEMEKNAQTIETLKQDWESERELCLENLRQELSLKHQSEMEGLQSQFQKELSEQKVELEKIFQAKHEAEVSLKNLEAQHQAAIKKLQEDLQSEHCQYLQDLEQKFREKEKAKELELETLQASYEDLKAQSQEEIRLLWSQLESMKTNREELNGSWDPVLAQASHLEELEHLRSGFAQQQQQERAQHESELEHLRVYFEKKLKDAEKTYQEDLTVFQQRLQEAREDSLESTEISSSCVLPEETSGREGKEPPDPLDLQLGQPKVQESLVEDCQVKLSKAEEKIQQMKEEFQKKEAEWELSREELKREAEERLASMFLELREKAESEKLSIISRFEHRESSMRHLQDQQAAQILDLERSLMEQQGHLRQLEQELTRDDLLPCSQCGQEPAMAQEEKNGALLREKEDCALQLLMAQNRFLEERKEIMEKFAKEQDAFLRDAQEKHNHELQLLQQGHQQQLLALRMELETKHRSELTEQLASSESRRQALLETHVAELQVKHNAEISALEKRHLSNLDELESCYVADVQTIRDEHQQALELLRAELEEQLQKKESCHREMLTQELENLKRQHAEELQSVRDSLRMEMSAQHIENGKGPAADLQGAHQQDPAMALHNEGHLLVEDGDAVLRSVDAEGLLHQAGPQELGDAHTVEMQKSQAELAKPQELQASQDQVAQVRDKVFLLNRELEECRAELEQLQQRRERENQEGTTLICMLRADLELAQGEGKALRDALRRLLDLFGDTLKAAVTLKSRISERAGLLLDHEDAADTSDARLAAAALGDMWSDEGLLEIDRTLPEGAETSSVCEISSHVCESFFISPENTLDCEQPIRRVYQSLSTAVEGLLEMALDSSKQLEEARQLHRCVEREFRHRNEEMAQAMQKQQELLERLREESAAKDRLALELHTAKGLLEGFKVEKVDLQEALGKKEESEQQLILELEDLRKQLQQAARELLTLKEEKSVLWNQKETLTNEAKEREAALQEEVESLTRVQWESRKQSEKDRATLLSQMRVLESELEDQLVQHRGCAQLAEEVATLKQQLAALDKHLRSQRQFMDDQAAEREHEREEFQQEIQRLEGQLRQAARPRPPGPRDSQCVQLDEEVELLQEKLREKLDGFNELVIKKDFADQQLLIQEEEIKRLEETNASIQRQMVQLQEELEKQKKSMEELKEKEILKQENMGDLLLTTVSRSGLDEAGCPMLPQGSSSRGPEAQPDVTERALLQHENEVVHRRNSEIDELKSLIENLQENQRQLQKDKAEEIEQLHEVIEKLQSELSLMGPKVHEVSDPQAGSLHSELACLRGEGLGGQALRSELQAAQAAKEVFGQLLANQAHGHSQALEALQQRLQDAEEVAARHLAELEHCVALREAEVEAMASQIQEFAATLKAKEAIIEQRDLEIDAVNKWKVSHSLELEAILLALAHFRHALEQQTCATPDEPPELRQLRVQCARLSHQLQVLYRPFLKCRMQLDQHQPHVASIGCANPCADDELEQEGVSNRLALAPHSLAAQAKEELEDCPLGKANLMAQVRQLQEELDHRVHSVASRDTNSETCKLQQPNLSENGPRNHCCNGEESKPSPPDDVLNIAKTTWDVIDIIKNQDLLVQVEMPDFPTQEKLTSQGGPFSSQASGHSGSLLPEEAAEPQQDPVRALDLSSWSSPEVVRKDPSLEPQHSLPLTPGVGTVSLHSVDISPDWTDPLLQADVSGLLCYPGKSASGQAPLWAVAPSAGKHHAERTATEKDVEDFIVTSFDSQELLTSPSHELARRSDGSRKSDGPDIAMMLTLGSEGSETPTTDLVAAAAAAVPFSRRFVQSPGAMKEKEIHAKQMKALLQMVFDESHQILALSESQDPSSALNKGEPRDPLDGFPRDSQALSEVTTDKGEKESLETHLTWSEELLRAIQEVFAREQEKAELQPRPYGSNLGDYNSLVQRLEKVIQEQGDPQKVQDHLCLSDRSSLLAEIQALRAQLRMTHLQNQEKLQQLCAALTSTEARGSQREHQLRRQVELLAYKVEQEKCIANELQKTLSKEQETASDVRKRLVVEQNAVQDLKSELHACKQENTSLLESLDKVQQEVLRLRAVLDGKEKELKVVLEELESERGKGQALQAQQEEQQLRYLQREGQSSRALEELKLSLEKQLAQNNQLCVALKHERAAKDNLQKELQIEASRCEALLAQEKGQLSELQKSLEAERSRSLELSEALQHERLLTEQLSRNSQEACARQETQVQHALLRKLKAEKTRALELEAMLEKVQKQAAHTQQQLEAQAQERCVELRREKERELEIQRQRDEHKIEQLQRLVRELRWKEEVSGGNGPCRGSPGRGSLERDQFQEQQQELEKIRQQLLCAAGLLTSFTNHTVDRTIKDWTSSNEKAVSSLMRTLEELKSELSMPTSFQKKMTAELQVQLMNELLSDNDALTKAVGMATREKAELCRTVSRLEKTLKHHTQKGCVLNRQSKSSLKQDGTDLQSSLRHSDPEWHSQTTSGDTNTCNIKMEKLYLHYLRAESFRKALIYQKKYLLLLIGGFQDSEQETLSMIAHLGVFPSKADKKITMSRPFTKFRTAVRVVIAVLRLRFLVKKWQEVDRKGALVHPKSTRHGHRTSQRQRSPSGPRASLPTRDTSSGPTKASRHSPRSAAAGSPGKERSTSTPSSRLERSLTASQDPEHSLTEYIHHLEMIQQRLGGLPPDSTQKSCHQKIKQ 50E66D93F7E4E283 nudc_mouse Nuclear migration protein nudC Nuclear distribution protein C homolog Silica-induced gene 92 protein Nudc protein protein mouse Mus musculus Mouse MGGEQEEERFDGMLLAMAQQHEGGVQELVNTFFSFLRRKTDFFIGGEEGMAEKLITQTFNHHNQLAQKARREKRARQETERREKAERAARLAKEAKAETPGPQIKELTDEEAERLQLEIDQKKDAEDQEAQLKNGSLDSPGKQDAEDEEDEEDEKDKGKLKPNLGNGADLPNYRWTQTLAELDLAVPFRVSFRLKGKDVVVDIQRRHLRVGLKGQPPVVDGELYNEVKVEESSWLIEDGKVVTVHLEKINKMEWWNRLVTSDPEINTKKINPENSKLSDLDSETRSMVEKMMYDQRQKSMGLPTSDEQKKQEILKKFMDQHPEMDFSKAKFN 83F30F2D8E013E3A tcpa_mouse T-complex protein 1 subunit alpha CCT-alpha Tailless complex polypeptide 1B Tcp1 Ccta Cct1 Tailless complex polypeptide 1A protein protein mouse Mus musculus Mouse MEGPLSVFGDRSTGEAVRSQNVMAAASIANIVKSSFGPVGLDKMLVDDIGDVTITNDGATILKLLEVEHPAAKVLCELADLQDKEVGDGTTSVVIIAAELLKNADELVKQKIHPTSVISGYRLACKEAVRYINENLIINTDELGRDCLINAAKTSMSSKIIGINGDYFANMVVDAVLAVKYTDARGQPRYPVNSVNILKAHGRSQIESMLINGYALNCVVGSQGMPKRIVNAKIACLDFSLQKTKMKLGVQVVITDPEKLDQIRQRESDITKERIQKILATGANVILTTGGIDDMYLKYFVEAGAMAVRRVLKRDLKHVAKASGASILSTLANLEGEETFEVTMLGQAEEVVQERICDDELILIKNTKARTSASIILRGANDFMCDEMERSLHDALCVVKRVLELKSVVPGGGAVEAALSIYLENYATSMGSREQLAIAEFARSLLVIPNTLAVNAAQDSTDLVAKLRAFHNEAQVNPERKNLKWIGLDLVHGKPRDNKQAGVFEPTIVKVKSLKFATEAAITILRIDDLIKLHPESKDDKHGSYENAVHSGALDD DIP protein P11983 original sequence version: 108 48F2387BE0F909A4 lis1_human Platelet-activating factor acetylhydrolase IB subunit beta PAF acetylhydrolase 45 kDa subunit PAF-AH alpha Lissencephaly-1 protein PAFAH1B1 LIS1 MDCR PAFAHA MDS protein protein human Homo sapiens Human MVLSQRQRDELNRAIADYLRSNGYEEAYSVFKKEAELDVNEELDKKYAGLLEKKWTSVIRLQKKVMELESKLNEAKEEFTSGGPLGQKRDPKEWIPRPPEKYALSGHRSPVTRVIFHPVFSVMVSASEDATIKVWDYETGDFERTLKGHTDSVQDISFDHSGKLLASCSADMTIKLWDFQGFECIRTMHGHDHNVSSVAIMPNGDHIVSASRDKTIKMWEVQTGYCVKTFTGHREWVRMVRPNQDGTLIASCSNDQTVRVWVVATKECKAELREHEHVVECISWAPESSYSSISEATGSETKKSGKPGPFLLSGSRDKTIKMWDVSTGMCLMTLVGHDNWVRGVLFHSGGKFILSCADDKTLRVWDYKNKRCMKTLNAHEHFVTSLDFHKTAPYVVTGSVDQTVKVWECR DIP protein P43034 original sequence version: 155 DIP protein P43034 original sequence version: 117 3AB68D2641BA31C9 lis1_cerae_protein protein protein chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey An amino acid sequence which is not represented in UniProt Similar to P63005 lis1_mouse Platelet-activating factor acetylhydrolase IB subunit beta PAF acetylhydrolase 45 kDa subunit PAF-AH alpha Lissencephaly-1 protein Pafah1b1 Lis1 Lis-1 Pafaha protein protein mouse Mus musculus Mouse MVLSQRQRDELNRAIADYLRSNGYEEAYSVFKKEAELDMNEELDKKYAGLLEKKWTSVIRLQKKVMELESKLNEAKEEFTSGGPLGQKRDPKEWIPRPPEKYALSGHRSPVTRVIFHPVFSVMVSASEDATIKVWDYETGDFERTLKGHTDSVQDISFDHSGKLLASCSADMTIKLWDFQGFECIRTMHGHDHNVSSVAIMPNGDHIVSASRDKTIKMWEVQTGYCVKTFTGHREWVRMVRPNQDGTLIASCSNDQTVRVWVVATKECKAELREHEHVVECISWAPESSYSSISEATGSETKKSGKPGPFLLSGSRDKTIKMWDVSTGMCLMTLVGHDNWVRGVLFHSGGKFILSCADDKTLRVWDYKNKRCMKTLNAHEHFVTSLDFHKTAPYVVTGSVDQTVKVWECR DIP protein P63005 original sequence version: 47 4DBF6A24A6B131CD slap1_mouse Src-like-adapter Sla Slap Slap1 Src-like-adapter protein 1 protein protein mouse Mus musculus Mouse MGNSMKSTSPPSERPLSSSEGLESDFLAVLTDYPSPDISPPIFRRGEKLRVISDEGGWWKAISLSTGRESYIPGICVARVYHGWLFEGLGRDKAEELLQLPDTKIGSFMIRESETKKGFYSLSVRHRQVKHYRIFRLPNNWYYISPRLTFQCLEDLVTHYSEVADGLCCVLTTPCLAQNIPAPTSHPSPCTSPGSPVTLRQKTFDWKRVSRLQEGSEGAENPLRVDESLFSYGLRESIASYLSLTGDDSSSFDRKKKSLSLMYTGSKRKSSFFSAPQYFED B347921656A74DA1 lis1-nudc 2 14 unspecified role unspecified role prey prey 16 unspecified role unspecified role bait bait physical association physical association Fig 1. We used a yeast two-hybrid screen to identify proteins that interact with LIS1 (Vojtek et al., 1993). Among the clones recovered, 31 encoded a previously identified LIS1 interactor, NudC (Morris et al., 1998a), while 14 encoded a new mouse protein homologous to the MP43, nudE, and RO11 genes. lis1-nude-1 2 11 unspecified role unspecified role prey prey 16 unspecified role unspecified role bait bait physical association physical association Fig 1. We used a yeast two-hybrid screen to identify proteins that interact with LIS1 (Vojtek et al., 1993). Among the clones recovered, 31 encoded a previously identified LIS1 interactor, NudC (Morris et al., 1998a), while 14 encoded a new mouse protein homologous to the MP43, nudE, and RO11 genes. Lis1 interacts with NudE (nde1). lis1-nude-2 7 11 unspecified role unspecified role bait bait n-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 16 unspecified role unspecified role prey prey physical association physical association Fig 2A. LIS1 specifically coimmunoprecipitates with myc-mNudE (Figure 2A), indicating that myc-mNudE associated with LIS1 when expressed in 293T cells. lis1-nude-4 6 16 unspecified role unspecified role bait bait P43034:p.His149Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain H R Wed May 02 16:02:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his149arg' Tue Jun 30 16:32:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His149Arg' P43034:p.Ser169Pro mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S P Wed May 02 20:08:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser169pro' Tue Jun 30 16:43:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser169Pro' 11 unspecified role unspecified role prey prey physical association physical association Fig 2C. Point mutations H149R and S169P were constructed in the LIS1 cDNA and then either fused to LexA or myc tagged. Both of the LIS1 missense mutations completely abolish LIS1-mNudE interaction in the two-hybrid assay (Figure 2C). The sites at amino acid positions 149 and 169 in the lis1 protein play an important role in binding to nude (nde1). lis1-nude-6 3 11 unspecified role unspecified role bait bait n-terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 16 unspecified role unspecified role prey prey n-terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag anti tag western anti tag western blot anti tag western n-term range n-terminal range n-term range n-term range n-terminal range n-term range P43034:p.His149Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain H R Wed May 02 18:17:06 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his149arg' Tue Jun 30 17:47:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His149Arg' P43034:p.Ser169Pro mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S P Wed May 02 16:01:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser169pro' Tue Jun 30 20:21:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser169Pro' physical association physical association Fig 2D. Furthermore, the two mutated LIS1 proteins failed to bind mNudE by coimmunoprecipitation (Figure 2D). Although others have claimed that the H149R and other LIS1 mutations are unstable (Sapir et al., 1999b), in our hands, the LIS1 missense mutations were expressed at levels comparable to the wild-type protein (Figure 2D). Lis1 interacts with NudE (nde1) and the two important amino acid sites at positions 149 and 169 are required for this interaction. nude-pcnt 2 13 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association Fig. 5A nude-cenpf 2 10 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association Fig. 6A nude-cep110 2 9 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association nude-gtubulin 4 12 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag anti tag western anti tag western blot anti tag western c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Fig 6A. We further examined possible physical interactions between mNudE, LIS1, and g-tubulin by transfecting 293T cells with GFP-tagged mNudE or GFP-tagged LIS1. Immunoprecipitation was performed with a g-tubulin monoclonal antibody and analyzed by nonreducing SDS-PAGE to avoid the interference of IgG with g-tubulin signals (48 kDa) on the immunoblot. The g-tubulin antibody selectively brought down full-length GFP-mNudE, or mNudE-GFP, but not GFP-LIS1 or the GFP-DN (Figure 6A). This suggests that mNudE forms a complex with g-tubulin in vivo and that LIS1 may be recruited to the centrosome via an interaction with mNudE, rather than associating with the g-tubulin complex directly. NudE (nde1) interacts with gamma-tubulin but not with Lis1. nude-tcp1 2 15 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association nude-slap 2 19 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association Fig 6A nude-lc8 2 11 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Two-hybrid interactions of mNudE with multiple centrosomal proteins. In a two-hybrid screen using the LexA-mNudE as the bait, clones encode partial or full-length sequence of pericentrin (Peri.), mitosin, CEP110, TCP-1, SLAP, and the 8 kDa light chain of cytoplasmic dynein (LC8) were identified. Fig. 6A nude-lc8-1 6 8 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association Fig 6B. (B) Two-hybrid interactions of mNudE with multiple centrosomal proteins. In a two-hybrid screen using the LexA-mNudE as the bait, clones encode partial or full-length sequence of pericentrin (Peri.), mitosin, CEP110, TCP-1, SLAP, and the 8 kDa light chain of cytoplasmic dynein (LC8) were identified. Plasmids from the two-hybrid positive clones were recovered and used to retransform L40 strain with LexA-mNudE or LexA alone. Two individual transformants were patched and assayed on a filter papers. Positive interactions are indicated by the blue b-galactosidase product. nude-tcp1-1 6 11 unspecified role unspecified role bait bait 15 unspecified role unspecified role prey prey physical association physical association Fig 6B. (B) Two-hybrid interactions of mNudE with multiple centrosomal proteins. In a two-hybrid screen using the LexA-mNudE as the bait, clones encode partial or full-length sequence of pericentrin (Peri.), mitosin, CEP110, TCP-1, SLAP, and the 8 kDa light chain of cytoplasmic dynein (LC8) were identified. Plasmids from the two-hybrid positive clones were recovered and used to retransform L40 strain with LexA-mNudE or LexA alone. Two individual transformants were patched and assayed on a filter papers. Positive interactions are indicated by the blue b-galactosidase product. nude-pcnt-1 6 13 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association Fig 6B. (B) Two-hybrid interactions of mNudE with multiple centrosomal proteins. In a two-hybrid screen using the LexA-mNudE as the bait, clones encode partial or full-length sequence of pericentrin (Peri.), mitosin, CEP110, TCP-1, SLAP, and the 8 kDa light chain of cytoplasmic dynein (LC8) were identified. Plasmids from the two-hybrid positive clones were recovered and used to retransform L40 strain with LexA-mNudE or LexA alone. Two individual transformants were patched and assayed on a filter papers. Positive interactions are indicated by the blue b-galactosidase product. nude-slap-1 6 11 unspecified role unspecified role bait bait 19 unspecified role unspecified role prey prey physical association physical association Fig 6B. (B) Two-hybrid interactions of mNudE with multiple centrosomal proteins. In a two-hybrid screen using the LexA-mNudE as the bait, clones encode partial or full-length sequence of pericentrin (Peri.), mitosin, CEP110, TCP-1, SLAP, and the 8 kDa light chain of cytoplasmic dynein (LC8) were identified. Plasmids from the two-hybrid positive clones were recovered and used to retransform L40 strain with LexA-mNudE or LexA alone. Two individual transformants were patched and assayed on a filter papers. Positive interactions are indicated by the blue b-galactosidase product. nude-cep110-1 6 9 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association Fig 6B. (B) Two-hybrid interactions of mNudE with multiple centrosomal proteins. In a two-hybrid screen using the LexA-mNudE as the bait, clones encode partial or full-length sequence of pericentrin (Peri.), mitosin, CEP110, TCP-1, SLAP, and the 8 kDa light chain of cytoplasmic dynein (LC8) were identified. Plasmids from the two-hybrid positive clones were recovered and used to retransform L40 strain with LexA-mNudE or LexA alone. Two individual transformants were patched and assayed on a filter papers. Positive interactions are indicated by the blue b-galactosidase product. nude-cenpf-1 6 10 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association Fig 6B. (B) Two-hybrid interactions of mNudE with multiple centrosomal proteins. In a two-hybrid screen using the LexA-mNudE as the bait, clones encode partial or full-length sequence of pericentrin (Peri.), mitosin, CEP110, TCP-1, SLAP, and the 8 kDa light chain of cytoplasmic dynein (LC8) were identified. Plasmids from the two-hybrid positive clones were recovered and used to retransform L40 strain with LexA-mNudE or LexA alone. Two individual transformants were patched and assayed on a filter papers. Positive interactions are indicated by the blue b-galactosidase product. lis1-nude-7 3 16 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role bait bait 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. Furthermore, the two mutated LIS1 proteins failed to bind mNudE by coimmunoprecipitation (Figure 2D). Although others have claimed that the H149R and other LIS1 mutations are unstable (Sapir et al., 1999b), in our hands, the LIS1 missense mutations were expressed at levels comparable to the wild-type protein (Figure 2D). lis1-nude-8 1 11 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 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) 18 unspecified role unspecified role prey prey direct interaction direct interaction Fig 2B. Binding of LIS1 to GST-mNudE. GST fusion proteins of full-length mNudE (GST-mNudE) lis1-nude-9 5 17 unspecified role unspecified role neutral component neutral component 11 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization Immunostaining also demonstrated that LIS1 is concentrated at the centrosome of interphase cells and costained with mNudE ( Figure 5 5J-5L).