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/ Cenp-F links kinetochores to Ndel1/Nde1/Lis1/dynein microtubule motor complexes. Cenp-F links kinetochores to Ndel1/Nde1/Lis1/dynein microtubule motor complexes. Curr. Biol. (0960-9822) 2007 imex curation Vergnolle MA., Taylor SS. Apoptosis - Interactions involving proteins with a function related to apoptosis stephen.taylor@manchester.ac.uk 23-Dec-2009: Contacted by JYOTI. A HeLa derived cDNA library was used. davet Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Vergnolle MA., Taylor SS. Apoptosis - Interactions involving proteins with a function related to apoptosis stephen.taylor@manchester.ac.uk Accepted 2009-NOV-8 by DAVET. 23-Dec-2009: Contacted by JYOTI. A HeLa derived cDNA library was used. Curr. Biol. (0960-9822) 2007 davet Only protein-protein interactions imex curation Cenp-F links kinetochores to Ndel1/Nde1/Lis1/dynein microtubule motor complexes. Cenp-F links kinetochores to Ndel1/Nde1/Lis1/dynein microtubule motor complexes. Curr. Biol. (0960-9822) 2007 imex curation Vergnolle MA., Taylor SS. Apoptosis - Interactions involving proteins with a function related to apoptosis stephen.taylor@manchester.ac.uk 23-Dec-2009: Contacted by JYOTI. A HeLa derived cDNA library was used. davet 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 Vergnolle MA., Taylor SS. Apoptosis - Interactions involving proteins with a function related to apoptosis stephen.taylor@manchester.ac.uk Accepted 2009-NOV-8 by DAVET. 23-Dec-2009: Contacted by JYOTI. A HeLa derived cDNA library was used. Curr. Biol. (0960-9822) 2007 davet Only protein-protein interactions imex curation Cenp-F links kinetochores to Ndel1/Nde1/Lis1/dynein microtubule motor complexes. Cenp-F links kinetochores to Ndel1/Nde1/Lis1/dynein microtubule motor complexes. Curr. Biol. (0960-9822) 2007 imex curation Vergnolle MA., Taylor SS. Apoptosis - Interactions involving proteins with a function related to apoptosis stephen.taylor@manchester.ac.uk 23-Dec-2009: Contacted by JYOTI. A HeLa derived cDNA library was used. davet 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 Vergnolle MA., Taylor SS. Apoptosis - Interactions involving proteins with a function related to apoptosis stephen.taylor@manchester.ac.uk Accepted 2009-NOV-8 by DAVET. 23-Dec-2009: Contacted by JYOTI. A HeLa derived cDNA library was used. Curr. Biol. (0960-9822) 2007 davet Only protein-protein interactions imex curation Cenp-F links kinetochores to Ndel1/Nde1/Lis1/dynein microtubule motor complexes. Cenp-F links kinetochores to Ndel1/Nde1/Lis1/dynein microtubule motor complexes. Curr. Biol. (0960-9822) 2007 imex curation Vergnolle MA., Taylor SS. Apoptosis - Interactions involving proteins with a function related to apoptosis stephen.taylor@manchester.ac.uk 23-Dec-2009: Contacted by JYOTI. A HeLa derived cDNA library was used. davet Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 fluorescence fluorescence technology fluorescence immunostaining immunostaining Vergnolle MA., Taylor SS. Apoptosis - Interactions involving proteins with a function related to apoptosis stephen.taylor@manchester.ac.uk Accepted 2009-NOV-8 by DAVET. 23-Dec-2009: Contacted by JYOTI. A HeLa derived cDNA library was used. Curr. Biol. (0960-9822) 2007 davet Only protein-protein interactions imex curation nde1_human Nuclear distribution protein nudE homolog 1 NDE1 NUDE protein protein human Homo sapiens Human MEDSGKTFSSEEEEANYWKDLAMTYKQRAENTQEELREFQEGSREYEAELETQLQQIETRNRDLLSENNRLRMELETIKEKFEVQHSEGYRQISALEDDLAQTKAIKDQLQKYIRELEQANDDLERAKRATIMSLEDFEQRLNQAIERNAFLESELDEKENLLESVQRLKDEARDLRQELAVQQKQEKPRTPMPSSVEAERTDTAVQATGSVPSTPIAHRGPSSSLNTPGSFRRGLDDSTGGTPLTPAARISALNIVGDLLRKVGALESKLASCRNLVYDQSPNRTGGPASGRSSKNRDGGERRPSSTSVPLGDKGLDTSCRWLSKSTTRSSSSC FC0BDD8BB90324B8 cenpf_human Centromere protein F CENPF Kinetochore protein CENPF Mitosin AH antigen protein protein human Homo sapiens Human MSWALEEWKEGLPTRALQKIQELEGQLDKLKKEKQQRQFQLDSLEAALQKQKQKVENEKTEGTNLKRENQRLMEICESLEKTKQKISHELQVKESQVNFQEGQLNSGKKQIEKLEQELKRCKSELERSQQAAQSADVSLNPCNTPQKIFTTPLTPSQYYSGSKYEDLKEKYNKEVEERKRLEAEVKALQAKKASQTLPQATMNHRDIARHQASSSVFSWQQEKTPSHLSSNSQRTPIRRDFSASYFSGEQEVTPSRSTLQIGKRDANSSFFDNSSSPHLLDQLKAQNQELRNKINELELRLQGHEKEMKGQVNKFQELQLQLEKAKVELIEKEKVLNKCRDELVRTTAQYDQASTKYTALEQKLKKLTEDLSCQRQNAESARCSLEQKIKEKEKEFQEELSRQQRSFQTLDQECIQMKARLTQELQQAKNMHNVLQAELDKLTSVKQQLENNLEEFKQKLCRAEQAFQASQIKENELRRSMEEMKKENNLLKSHSEQKAREVCHLEAELKNIKQCLNQSQNFAEEMKAKNTSQETMLRDLQEKINQQENSLTLEKLKLAVADLEKQRDCSQDLLKKREHHIEQLNDKLSKTEKESKALLSALELKKKEYEELKEEKTLFSCWKSENEKLLTQMESEKENLQSKINHLETCLKTQQIKSHEYNERVRTLEMDRENLSVEIRNLHNVLDSKSVEVETQKLAYMELQQKAEFSDQKHQKEIENMCLKTSQLTGQVEDLEHKLQLLSNEIMDKDRCYQDLHAEYESLRDLLKSKDASLVTNEDHQRSLLAFDQQPAMHHSFANIIGEQGSMPSERSECRLEADQSPKNSAILQNRVDSLEFSLESQKQMNSDLQKQCEELVQIKGEIEENLMKAEQMHQSFVAETSQRISKLQEDTSAHQNVVAETLSALENKEKELQLLNDKVETEQAEIQELKKSNHLLEDSLKELQLLSETLSLEKKEMSSIISLNKREIEELTQENGTLKEINASLNQEKMNLIQKSESFANYIDEREKSISELSDQYKQEKLILLQRCEETGNAYEDLSQKYKAAQEKNSKLECLLNECTSLCENRKNELEQLKEAFAKEHQEFLTKLAFAEERNQNLMLELETVQQALRSEMTDNQNNSKSEAGGLKQEIMTLKEEQNKMQKEVNDLLQENEQLMKVMKTKHECQNLESEPIRNSVKERESERNQCNFKPQMDLEVKEISLDSYNAQLVQLEAMLRNKELKLQESEKEKECLQHELQTIRGDLETSNLQDMQSQEISGLKDCEIDAEEKYISGPHELSTSQNDNAHLQCSLQTTMNKLNELEKICEILQAEKYELVTELNDSRSECITATRKMAEEVGKLLNEVKILNDDSGLLHGELVEDIPGGEFGEQPNEQHPVSLAPLDESNSYEHLTLSDKEVQMHFAELQEKFLSLQSEHKILHDQHCQMSSKMSELQTYVDSLKAENLVLSTNLRNFQGDLVKEMQLGLEEGLVPSLSSSCVPDSSSLSSLGDSSFYRALLEQTGDMSLLSNLEGAVSANQCSVDEVFCSSLQEENLTRKETPSAPAKGVEELESLCEVYRQSLEKLEEKMESQGIMKNKEIQELEQLLSSERQELDCLRKQYLSENEQWQQKLTSVTLEMESKLAAEKKQTEQLSLELEVARLQLQGLDLSSRSLLGIDTEDAIQGRNESCDISKEHTSETTERTPKHDVHQICDKDAQQDLNLDIEKITETGAVKPTGECSGEQSPDTNYEPPGEDKTQGSSECISELSFSGPNALVPMDFLGNQEDIHNLQLRVKETSNENLRLLHVIEDRDRKVESLLNEMKELDSKLHLQEVQLMTKIEACIELEKIVGELKKENSDLSEKLEYFSCDHQELLQRVETSEGLNSDLEMHADKSSREDIGDNVAKVNDSWKERFLDVENELSRIRSEKASIEHEALYLEADLEVVQTEKLCLEKDNENKQKVIVCLEEELSVVTSERNQLRGELDTMSKKTTALDQLSEKMKEKTQELESHQSECLHCIQVAEAEVKEKTELLQTLSSDVSELLKDKTHLQEKLQSLEKDSQALSLTKCELENQIAQLNKEKELLVKESESLQARLSESDYEKLNVSKALEAALVEKGEFALRLSSTQEEVHQLRRGIEKLRVRIEADEKKQLHIAEKLKERERENDSLKDKVENLERELQMSEENQELVILDAENSKAEVETLKTQIEEMARSLKVFELDLVTLRSEKENLTKQIQEKQGQLSELDKLLSSFKSLLEEKEQAEIQIKEESKTAVEMLQNQLKELNEAVAALCGDQEIMKATEQSLDPPIEEEHQLRNSIEKLRARLEADEKKQLCVLQQLKESEHHADLLKGRVENLERELEIARTNQEHAALEAENSKGEVETLKAKIEGMTQSLRGLELDVVTIRSEKENLTNELQKEQERISELEIINSSFENILQEKEQEKVQMKEKSSTAMEMLQTQLKELNERVAALHNDQEACKAKEQNLSSQVECLELEKAQLLQGLDEAKNNYIVLQSSVNGLIQEVEDGKQKLEKKDEEISRLKNQIQDQEQLVSKLSQVEGEHQLWKEQNLELRNLTVELEQKIQVLQSKNASLQDTLEVLQSSYKNLENELELTKMDKMSFVEKVNKMTAKETELQREMHEMAQKTAELQEELSGEKNRLAGELQLLLEEIKSSKDQLKELTLENSELKKSLDCMHKDQVEKEGKVREEIAEYQLRLHEAEKKHQALLLDTNKQYEVEIQTYREKLTSKEECLSSQKLEIDLLKSSKEELNNSLKATTQILEELKKTKMDNLKYVNQLKKENERAQGKMKLLIKSCKQLEEEKEILQKELSQLQAAQEKQKTGTVMDTKVDELTTEIKELKETLEEKTKEADEYLDKYCSLLISHEKLEKAKEMLETQVAHLCSQQSKQDSRGSPLLGPVVPGPSPIPSVTEKRLSSGQNKASGKRQRSSGIWENGRGPTPATPESFSKKSKKAVMSGIHPAEDTEGTEFEPEGLPEVVKKGFADIPTGKTSPYILRRTTMATRTSPRLAAQKLALSPLSLGKENLAESSKPTAGGSRSQKVKVAQRSPVDSGTILREPTTKSVPVNNLPERSPTDSPREGLRVKRGRLVPSPKAGLESNGSENCKVQ 1FC81972F947B375 ndel1_human Nuclear distribution protein nudE-like 1 NDEL1 Mitosin-associated protein 1 EOPA MITAP1 NUDEL protein protein human Homo sapiens Human MDGEDIPDFSSLKEETAYWKELSLKYKQSFQEARDELVEFQEGSRELEAELEAQLVQAEQRNRDLQADNQRLKYEVEALKEKLEHQYAQSYKQVSVLEDDLSQTRAIKEQLHKYVRELEQANDDLERAKRATIVSLEDFEQRLNQAIERNAFLESELDEKESLLVSVQRLKDEARDLRQELAVRERQQEVTRKSAPSSPTLDCEKMDSAVQASLSLPATPVGKGTENTFPSPKAIPNGFGTSPLTPSARISALNIVGDLLRKVGALESKLAACRNFAKDQASRKSYISGNVNCGVLNGNGTKFSRSGHTSFFDKGAVNGFDPAPPPPGLGSSRPSSAPGMLPLSV DIP protein Q9GZM8 original sequence version: 36 DIP protein Q9GZM8 original sequence version: 65 DIP protein Q9GZM8 original sequence version: 78 E91EA053FCA315D7 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 bub1_human Mitotic checkpoint serine/threonine-protein kinase BUB1 BUB1A BUB1 BUB1L protein protein human Homo sapiens Human MDTPENVLQMLEAHMQSYKGNDPLGEWERYIQWVEENFPENKEYLITLLEHLMKEFLDKKKYHNDPRFISYCLKFAEYNSDLHQFFEFLYNHGIGTLSSPLYIAWAGHLEAQGELQHASAVLQRGIQNQAEPREFLQQQYRLFQTRLTETHLPAQARTSEPLHNVQVLNQMITSKSNPGNNMACISKNQGSELSGVISSACDKESNMERRVITISKSEYSVHSSLASKVDVEQVVMYCKEKLIRGESEFSFEELRAQKYNQRRKHEQWVNEDRHYMKRKEANAFEEQLLKQKMDELHKKLHQVVETSHEDLPASQERSEVNPARMGPSVGSQQELRAPCLPVTYQQTPVNMEKNPREAPPVVPPLANAISAALVSPATSQSIAPPVPLKAQTVTDSMFAVASKDAGCVNKSTHEFKPQSGAEIKEGCETHKVANTSSFHTTPNTSLGMVQATPSKVQPSPTVHTKEALGFIMNMFQAPTLPDISDDKDEWQSLDQNEDAFEAQFQKNVRSSGAWGVNKIISSLSSAFHVFEDGNKENYGLPQPKNKPTGARTFGERSVSRLPSKPKEEVPHAEEFLDDSTVWGIRCNKTLAPSPKSPGDFTSAAQLASTPFHKLPVESVHILEDKENVVAKQCTQATLDSCEENMVVPSRDGKFSPIQEKSPKQALSSHMYSASLLRLSQPAAGGVLTCEAELGVEACRLTDTDAAIAEDPPDAIAGLQAEWMQMSSLGTVDAPNFIVGNPWDDKLIFKLLSGLSKPVSSYPNTFEWQCKLPAIKPKTEFQLGSKLVYVHHLLGEGAFAQVYEATQGDLNDAKNKQKFVLKVQKPANPWEFYIGTQLMERLKPSMQHMFMKFYSAHLFQNGSVLVGELYSYGTLLNAINLYKNTPEKVMPQGLVISFAMRMLYMIEQVHDCEIIHGDIKPDNFILGNGFLEQDDEDDLSAGLALIDLGQSIDMKLFPKGTIFTAKCETSGFQCVEMLSNKPWNYQIDYFGVAATVYCMLFGTYMKVKNEGGECKPEGLFRRLPHLDMWNEFFHVMLNIPDCHHLPSLDLLRQKLKKVFQQHYTNKIRALRNRLIVLLLECKRSRK DIP protein O43683 original sequence version: 94 DIP protein O43683 original sequence version: 154 38AE5E1F88C53BDC cenpf-ndel1-6 2 7 unspecified role unspecified role prey prey c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 6 unspecified role unspecified role bait bait c-terminus sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association Fig S1A, B and C. To define Cenp-F's role in kinetochore-microtubule interactions (KT-MT), we screened a yeast two-hybrid (Y2H) library by using a C-terminal Cenp-F fragment (fragment a, Figure S1A in the Supplemental Data available online). This identified Ndel1, one of two human NudE-related proteins, the other being Nde1 (see [8]). In addition to full-length Ndel1, the screen yielded C-terminal Ndel1 fragments, suggesting that Ndel1 binds Cenp-F via its NudC domain (Figure S1C). To identify the minimal domain of Cenp-F capable of interacting with Ndel1, fragment a was further fragmented, thereby identifying a 100 amino acid region as containing the Ndel1-binding domain (Figures S1A and S1B). The c-terminus of cenpf was able to interact with Ndel1. cenpf-ndel1-7 3 6 unspecified role unspecified role bait bait c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role prey prey c-terminus sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association Fig S1A, B and C. To define Cenp-F's role in kinetochore-microtubule interactions (KT-MT), we screened a yeast two-hybrid (Y2H) library by using a C-terminal Cenp-F fragment (fragment a, Figure S1A in the Supplemental Data available online). This identified Ndel1, one of two human NudE-related proteins, the other being Nde1 (see [8]). In addition to full-length Ndel1, the screen yielded C-terminal Ndel1 fragments, suggesting that Ndel1 binds Cenp-F via its NudC domain (Figure S1C). To identify the minimal domain of Cenp-F capable of interacting with Ndel1, fragment a was further fragmented, thereby identifying a 100 amino acid region as containing the Ndel1-binding domain (Figures S1A and S1B). The c-terminus of cenpf interacts with the c-terminus of Ndel1. ndel1-bub1 4 9 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization Fig S1D and Fig 1D. Having identified Ndel1 as a Cenp-F-binding partner, we asked whether Ndel1 localizes to KTs. During mitosis, transfected full-length Myc-Ndel1 colocalized with Bub1 at KTs (Figure S1D). We next asked whether KT localization of Ndel1 was dependent on Cenp-F. Importantly, Cenp-F RNAi abolished KT localization of Ndel1 (Figure 1D). Only the full-length and c-terminus of Ndel1 were able to colocalize with Bub1 at the kinetochore. The localization of Ndel1 was dependant on Cenpf. cenpf-ndel1-8 1 7 unspecified role unspecified role prey prey 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 unspecified role unspecified role bait bait 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 c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association Fig 1A. To confirm that the Y2H data reflected a physical interaction, we incubated GST-Ndel1 in HeLa extracts expressing GFP-tagged Cenp-F fragments. Importantly, Cenp-F fragment d8 bound GST-Ndel1 (Figure 1A). Consistent with Cenp-F binding the C-terminal domain of Ndel1, fragment d8 did not bind GST fused to the N terminus of Ndel1 (Figure 1A). The c-terminus of cenpf interacts with full-length Ndel1. cenpf-nde1-6 1 5 unspecified role unspecified role prey prey 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 unspecified role unspecified role bait bait 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 c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association Fig 3E. In addition, Cenp-F fragment d8 bound GST-Nde1 in the in vitro binding assay (Figure 3E). Thus, Cenp-F binds and targets Nde1 to KTs. The c-terminus of cenpf interacts with full-length Nde1. ndel1-cenpf 4 6 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 c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 40 43 colocalization colocalization Fig S2B. To determine whether Ndel1 targets KTs via a direct interaction with Cenp-F, we expressed GFP-tagged Cenp-F fragments with different abilities to bind Ndel1 and/or target the KT. Importantly, fragment d8, which binds Ndel1 but does not localize to KTs, titrated Ndel1 away from KTs (Figure S2B). By contrast, fragment a, which binds Ndel1 and localizes to KTs, and fragment c, which does not bind Ndel1 and does not localize to KTs, did not titrate Ndel1 away from KTs (Figure S2B). Cenp-F targets Ndel1 to kinetochores via a direct interaction. lis1-cenpf-1 4 8 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig S3. While repression of Cenp-F dislodged Myc-Lis1 from KTs, Myc-Lis1 remained KT bound after inhibition of either Ndel1 or Nde1 (Figure S3). Cenp-F targets Lis1 to kinetochores.