[email protected]) ([email protected]; correspondence for *Authors 2705, CHUQ, du Recherche Que de T3-51, Centre Laurier, Health, Boulevard Child and Health Perinatal cecs xodBoksUiest,Ofr X B,UK. 0BP, OX3 Oxford University, Brookes Oxford Sciences, h yai rti n1–akntcoerendezvous Ghongane a Priyanka – Knl1 dynamic The COMMENTARY ß n1fnto aebe mlctdi eoeinstability, genome 1 (Kiyomitsu in in disorders Defects implicated neurological 2012). and been al., microcephaly et have improperly leukemia, Espeut function 2011; or al., al., et unattached Knl1 Meadows et assembly 2007; of Rosenberg delays al., spindle presence 2011; et that (Kiyomitsu the the cascade in attached signaling inactivating onset conserved and anaphase a accurate (SAC), activating for and checkpoint required 2003), the both al., is et of Knl1, network, (Desai for mitosis 2009). formation KMN during segregation the Musacchio, the and of and subunit to (Przewloka Santaguida largest contributes hub 2009; attachments signaling and Glover, a (MT) as mitosis kinetochore–microtubule functions 2011; early Knl1 Stukenberg, 2012). during and Salmon, Tooley and Spc25] 2012; Ndc80 and Varma Spc24 Musacchio, the Nuf2, and and Ndc80), Nnf1), as (DeLuca the known and (also Nsl1 comprises a [Hec1 Dsn1, complex is also (Mis12, Blinkin, that complex and Mis12 assembly a fission AF15q14 network, macromolecular in (KMN) CASC5, ten-subunit Knl1–Mis12–Ndc80 Spc7 the as yeast, of humans component budding central in in Spc105 and as yeast, known also Knl1, Introduction Mitosis, Kinetochore, checkpoint motif, Mitotic MELT Knl1, BubR1, WORDS: KEY binding Knl1. by mediate activity that scaffolding interactions and Knl1 recognition cooperative characterize partner the that in complexity implicated structural low are explore of we the Finally, regions of B. the regulation Aurora how and and recruitment Bub3 BubR1, the Bub1, in role MELT SAC their SILK, RVSF, and motifs, the KI including and Knl1, protein in the present of motifs a the roles as interaction emerging protein present of the this We of platform. roles regulation discuss docking the versatile and we exquisite Knl1 Commentary, of organization the domain this overall ensures In phosphatases, which processes. and aforementioned kinases of protein interplay certain of the substrate the kinetochore is the to and have components SAC recruits reports Knl1 in functions organisms. Knl1 higher provided dictate recent and and that features processes structural these and of function details in molecular Knl1 number of (SAC) prominence the A confirmed checkpoint congression. assembly chromosome spindle kinetochore proper for evolutionarily assembly, required an is is that Q8NG31) protein scaffolding UniProt conserved CASC5, as known (also Knl1 ABSTRACT aut fHat n ieSine,Dprmn fBooia n Medical and Biological of Department Sciences, Life and Health of Faculty 04 ulse yTeCmayo ilgssLd|Junlo elSine(04 2,31–43doi:10.1242/jcs.149922 3415–3423 127, (2014) Science Cell of Journal | Ltd Biologists of Company The by Published 2014. ´ 1 e,Q 1 G,Canada. 4G2, G1V QC bec, ai Kapanidou Maria , 1 de Asghar Adeel , 2 Reproduction, 2 aieElowe Sabine , Bx1.Kl srqie o uoaBmdae hshrlto of phosphorylation Aurora-B-mediated is for itself required kinetochore is B the Knl1 and Aurora 1). MTs (Box between activity. attachments as B the emerged govern Aurora to 2013; also thought for has al., ‘switchboard’ Knl1 et 2013). central Varma DeLuca, 2012; the and al., Caldas et by Venkei reviewed 2008; 2009; al., et al., recent Cheeseman et 2003; of al., Schittenhelm a et number (Desai cells signaling, major a human cultured BubR1 from a and support and that yeasts, significant Bub1 in suggesting seen studies coordinate 2008), has to that al., is notion et Knl1 of the Cheeseman Cheeseman function 2011; with 2006; al., associated et al., Kiyomitsu and that et 2007; Bub1 al., to proteins in et checkpoint (Kiyomitsu similar Genin cells mitotic BubR1 2014; the in is of al., observed depleted expression et suppressed phenotype is Yang the Knl1 2011; Interestingly, which al., 2012). et al., Kiyomitsu et 2007; al., et npriua,rcn nihsit h ucinadrglto of regulation and Knl1. function a of the motifs this into advance protein-interacting by insights the mediated and recent emphasize, are particular, the We mitosis segregation. in that chromosome this of proper interactions during ensure In to C- the understanding protein Knl1 of 1). a (Fig. current view of and mechanistic fold our functions RWD protein–protein KI, discuss different the and the of we adopts MELT Commentary, characterization that including RVSF, domain and Knl1, SILK, terminal identification motifs in interaction recent regions emerged that have the activities platform functional activities. these to phosphatase coordinates docking and Knl1 a owing how kinase to as SAC as of functions Clues number Knl1 a important provided integrates how have reports into recent Kruse of 2012; insight number al., A et 2013). Suijkerbuijk al., 2010; et al., (indirectly et PP2A well (Liu and kinase as BubR1) (directly) PP1 through 2007), SAC phosphatases al., protein the the et of of (Kiyomitsu as recruitment BubR1 and kinetochore pseudokinase 2012) the and al., domain. Bub1 et in Espeut roles globular 2010; key al., C-terminal structural et has a (Welburn low MTs of of to binds regions presence Knl1 of the abundance and an complexity by characterized are the 2013). 2006; DeLuca, to primary al., and contributes Caldas et that 2010; (Kline the al., 2006; protein et attachments a Welburn al., kinetochore–MT Dsn1, Hec1, and of et 2010), stabilization (Cheeseman including al., et protein Alushin proteins, attachment kinetochore–MT kinetochore outer ltom h -emnlrgo fKl srsosbefrits for responsible is Knl1 of of region variety C-terminal overall docking A its multifactorial The a with as platform. 2013). Knl1 together define DeLuca, complexity, partners, structural protein low and al., and et Caldas motifs Pagliuca 2007; functional in al., reviewed et machinery Kiyomitsu 2009; congression 2008; chromosome al., proteins et and as well (Cheeseman SAC as the kinetochore itself, in of network number implicated KMN a the of within recruitment both the proteins, for machinery, important scaffolding is kinetochore the Knl1 of protein component central scaffolding a kinetochore As major a – Knl1 n1otoosso o mn cdsqec iiaiyand similarity sequence acid amino low show orthologs Knl1 2, anradtselegans Caenorhabditis n itrM Bolanos-Garcia M. Victor and * , rspiamelanogaster Drosophila 1, * 3415

Journal of Cell Science COMMENTARY 00 oebr ta. 01,a nyeta oneat the counteracts that 3416 and enzyme Knl1 an 2011), of al., et al., N-terminus (Liu et PP1 far phosphatase Rosenberg protein the 2010; of at binding direct located the (consensus mediate are RVSF and motifs (SG)ILK] RVxF) sequence [consensus SILK The motifs RVSF and SILK regions protein-interacting Knl1 reported. between been complex not al., protein has et stable Knl1 Wei a and 2006; for Hec1 al., evidence KMN et although reconstituted (Cheeseman patch 2007), a cooperatively act of basic to context conserved shown the highly In from 2010). a network MT- al., N-terminal includes et an through that (Welburn MTs to region orthologs bind Knl1 directly binding 2014). to R al., able residues et also (Petrovic are act acid Nsl1 complexes amino with Knl1, interact two the human directly the In encompassing kinetochore. that fragment the possible to Zwint a is Knl1 the and recruit it Because to Mis12 2004). 2013), cooperatively on al., al., depends et et Wang Knl1 (Varma 2005; of al., localization et kinetochore Kops the 2011; (Petrovic binding (Kiyomitsu al., for Zwint prometaphase et required in to is complex that binding (RZZ) protein 1904– Rod–Zwilch–Zw10 its kinetochore (residues a mediates 2010) Knl1 al., also of et al., humans) region al., in et C-terminal et The 2316 Petrovic (Cheeseman 2010; 1). al., complex (Fig. et Mis12 Petrovic 2014) with 2007; the al., interaction et of Kiyomitsu its 2006; component for required a is Nsl1, and localization kinetochore 09,atog h infcneo h neato ean unclear al., 2011). remains al., et interaction et the (Glatter (Foley of PP2A significance the phosphatase although protein 2009), of subunits regulatory hte hshtssta r erie yKl regulate B55- of SKA3 Knl1 partner question. binding open a by 2012). an as identified is al., recruited been SKA also et and has are (Chan KMN between activity that is interactions B networks al., phosphatases SKA et Aurora and Raaijmakers Whether by KMN 2009; regulated the al., between et negatively Daum association 2009; The al., 2009). al., stable et et kinetochore- maintaining Gaitanos Theis 2006; in and 2009; and al., implicated SKA2 et (Hanisch strongly spindle SKA1, interactions been kinetochore–MT by the has formed which with complex SKA3, and protein interact (SKA) Lampson directly associated to 2013; see shown longer Tanaka, no reviews 2013; thus recent are Wynne, 2011). and excellent Cheeseman, B and are (for Aurora B Funabiki to of phosphorylated accessible Aurora influence’ less efficiently spindle of of become of they ‘sphere targets centromere; the consequence kinetochore the a from outer displaced as the the occurs repair the or biorientation, that to tension incorrect stretch machinery of application kinetochore of kinetochore thus the Upon and interactions, result attachments. inappropriate spindle kinetochore–MT a the these as allowing of the centromere in stability results decreased the then Phosphorylation to attachments. centromere kinetochore–MT so-called the into proximity brought the the are at close causes that proteins that literature, B outer-kinetochore gradient of activity Aurora the phosphorylation radiating that outwardly is in an posits B generates model Aurora model, current by displacement attachments The kinetochore–MT complex. of regulation The kinetochore–MT of regulation attachments B Aurora 1. Box M ewr rtis(lhuhntKl tef aeas been also have itself) Knl1 not (although proteins network KMN .elegans C. hs Tbnigatvte aebeen have activities MT-binding these , b n B55- and 2096 –D d 2311 , usrts h ute lcdto fhwtekinetochore the how of elucidation further different upon The act potentially enzymes substrates. two is recruitment these PP2A, kinetochore PP2A and the PP1 mediates Kruse of 2013). 2012; Knl1 al., Although et 2013). al., Suijkerbuijk 2010; al., al., et mitosis et et Liu late 2011; Xu in al., through et substrates (Foley 2013; kinetochore attachments (Suijkerbuijk outer al., kinetochore–MT subunits of dephosphorylation et stabilizing regulatory Kruse for PP2A directly important 2012; of binds al., family which of et B56 BubR1, recruitment of the the recruitment mediates to indirectly the Knl1 through PP1, PP2A negatively 2011). cells al., to et Rosenberg addition in PP1 2014; 2011; al., In al., et et that Meadows that (Zhang 2012; al., BubR1 with et and London suggesting Bub1 compared of recruitment Knl1, kinetochore the regulates the wild-type at expressing N- the Bub1 BubR1 of of mutation levels increased the and in or results Knl1, site, of PP1-binding acids Knl1 of amino terminal deletion 300 agreement or that 150 reported In first recently and 2012). the laboratory al., Bub1 Nilsson the et of this, (Espeut removal with kinetochore al., timely the the et from ensure BubR1 (Liu might motif which RVSF silencing, and the feedback PP1 In a of between 2010). Through phosphorylation silencing interaction 2012). through SAC the al., inhibits Knl1 mediates et B that Espeut Aurora and 2011; mechanism, al., B et Aurora (Lesage of activity kinase ih lopooeArr iaeatvt Cla ta. 2013). al., et equivalent (Caldas Knl1 activity an kinase of B in regions Aurora other promote result that also suggesting not might activity, the with B does Aurora localization promotes of fragment B rescue turn Aurora Knl1 of N-terminal in rescue Yamagishi an Interestingly, which 2013; 2012). al., substrate event et al., Bub1 (Caldas et an B the Aurora of T120, of targeting centromere at phosphorylation H2A the histone facilitates N-terminus the Knl1 Work that indirectly, demonstrated below). of has occurs (see laboratory DeLuca Knl1 Bub1 the by of from regulation recruitment B Knl1-mediated Aurora through versus of antibodies facet phosphospecific second of B Fo to use Aurora targeted due for the used be the were [namely, might that but for activity readouts previous clear, the reason of not sensitivities is The different to studies 2013). these B al., between Aurora contrast disparity affect et significantly site not (Caldas PP1-binding in does and the autophosphorylation AAAA) of to Caldas that, (RVSF mutation Indeed, Knl1 2010), of al., 2010). shown et al., (Liu et observations have (Liu phosphorylation N- substrates enhance collaborators the to B to expected Aurora PP1 is of of Knl1 binding of of region loss somewhat terminal the 2013). is that diminished al., is considering et activity surprising, B resulting (Caldas Aurora thus that attachment observation kinetochore–MT Dsn1, The outer in and of defects Hec1 in including phosphorylation proteins, of Aurora-B-mediated depletion kinetochore or region abolishes N-terminal Knl1 a Knl1 of Deletion B. mechanism Aurora of the into is insights phosphatases regulation. important these kinetochore underlying provide of will selectivity achieved substrate and localization Cesmne l,20;Vegle l,21;Vegle al., et feature Vleugel 2012; conserved motif al., MELT et evolutionary the Vleugel an 2004; of al., units being sequence et multiple (Cheeseman a Knl1/Spc105 3). of 1, with (Figs presence in orthologs motif Knl1 the of MELT the regions Despite middle in the the found and of are N-terminal (M/I/L/V)-(E/D)-(L/M/I/V)-(T/S) copies of of consensus number A motifs MELT ao ucino n1budP1i orglt h activity the regulate to is PP1 Knl1-bound of function major A ora fCl cec 21)17 4532 doi:10.1242/jcs.149922 3415–3423 127, (2014) Science Cell of Journal .elegans C. rtrrsnneeeg rnfr(RT esr] A sensors]. 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Journal of Cell Science Bub1 N I COMMENTARY ii ET(rmrce l,21) h tutr fthe of located structure a are that define that The end residues 2013). Bub1 N-terminal phosphopeptides few the (ITC) al., a at synthetic that calorimetry et shows complex with titration (Primorac ternary MELT isothermal Bub3 mimic by of the supported the experiments notion in between a is affinity, directly of interaction moderate a that the has engage Bub3 phosphothreonine suggesting and not MELT thus phosphorylated motifs, the Bub3, do MELT with flank motif with interaction Bub3 MELT that of phosphorylated to interaction residues appear that residues the site Bub1 whereas in few the 2013; state, implicated as al., crystalline be 2) et the (Primorac (Box In motif 2A,B). repeats MELT Fig. WD40 phosphorylated the Bub3 a to the identified binds by motif) formed the GLEBS of is the side the as on protein region to (a well-conserved MELT referred Bub1 Bub3 synthetic of commonly a region to with motif structure Bub3-binding complex affinity crystal the in The and yeast high 2014). phosphopeptide budding al., with from et Bub3 Zhang and phosphorylated of and 2013; al., directly Structural that et bind (Primorac studies. demonstrated motifs recent has MELT more evidence in investigated biochemical been an has mount 2012). Shepperd al., to 2012; et Yamagishi al., 2012; failure al., et et (London and Knl1, response to checkpoint defects, proteins appropriate BUB congression the by of binding chromosome Knl1 attenuated of in phosphorylation results Mps1, the al., al., preventing et et Yamagishi Accordingly, 2012; (London al., 2012). kinetochore et Shepperd the 2013; al., to et subset Primorac the Bub3 2012; for a and required and Bub1 least is Mps1, of at Knl1 kinase recruitment of checkpoint or modification mitotic – posttranslational the motifs this of that target MELT the the is of – thereof 4 al., position et in residue Vleugel a 2012; is al., Knl1 et that suggests (Vleugel 2013). thus protein a and evolving shows species motifs rapidly MELT-like across or variation MELT of large number exact the 2013), i detail more in recru described Bub3 is instance, that For interactions. signaling the SAC of of regulation aspect PP1 important the recruit an in to Mps1, factor Knl1 key by of Knl1 a ability of be the repeats addition, to motif likely MELT is the Knl1 of phosphorylation of requires N-terminal kinetochore the on activities phosphatase scaffold. kinetochore and kinase a is Knl1 1. Fig. four-stranded b eietolo ein;namely, regions; loop two define eiusta r nae nteitrcinwt u3(Primorac Bub3 with interaction the in engaged are that residues 309 1– 244 ti o salse htpopoyaino h threonine the of phosphorylation that established now is It h usino o ETsqecsrcutBBproteins BUB recruit sequences MELT how of question The b n E and n Bub3 and 2 n N and (S/G)ILKRVSF b MT binding hipnpyial neatwt h u3rsde that residues Bub3 the with interact physically -hairpin PP1 B Aurora 249 315 γ –A –I b setlctdaoetepopoyae MELT phosphorylated the above located -sheet 317 251 b epciey.Aioai eiuso this of residues acid Amino respectively). , epciey.TeitrcinbtenBub1 between interaction The respectively). , A6– b MELT Bub3

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Journal of Cell Science COMMENTARY p15Kl ucin(htehl ta. 09.Further 2009). are 3418 motifs MELT al., repeating in that et showed cells, also (Shittenhelm analysis mammalian sequence and function an yeasts Spc105/Knl1 play in 2013). not al., situation et melanogaster might (Vleugel the flies Mps1 in sequence recruitment that Unlike BUB MELT threonine suggesting in role the the glutamate, important the by of or replaced where been aspartate phosphorylation has drosophilids, residues sequences repeat MELT acidic MELT-like in the the of identified of phosphoacceptor divergent been (Vleugel most The exist have 2013). do The al., variances et negatively motif. species-specific often although are MELT highly residues are charged, intervening the motif the MELT whereas the resemble of conserved, residues recent that threonine of A and presence modules methionine evolution. the repeating revealed across Knl1 apparent 19 human also of analysis is bioinformatics motif MELT core MELT SAC individual of the functionality the and to motifs. determine congression capacity to needed their chromosome be systematic will and BUB A binding, 2014). phosphorylation of Bub3 al., motif levels support et MELT higher Krenn of 2013; require analysis al., might et (Vleugel pairs activity chromatid the alignment supports sister that suggesting construct of congression, Knl1 MELT chromosome not localized N-terminal remaining but When this SAC 2013). the al., kinetochore, et the providing (Vleugel to present congression, are Knl1 or required of strictly motifs SAC not is the regions region chromosomal Knl1 for ectopic this motifs, to However, KI2 proteins mitosis. and Bub1 during KI1 recruit the to with able together are motif, MELT first the only hcpitpoen htcnanteW4 odaeBb and Bub3 are mitotic fold of WD40 Examples the ligands. contain protein Cdc20. that of docking proteins binding rigid checkpoint a the as serve for units platform repeat WD40 WD40- multi-protein structure. the of assembly blade where the 3D complexes, in the involved form usually are to proteins repeat repeat next seven- the of a three last first the with that domain, so interlock WD40 the circular acid a bladed called adopt tryptophan-aspartic to architecture together pair fold of blades solenoid The the motif name. the in structural hence (WD), terminating short often a residues, of consists anti-parallel repeat repeats four-stranded WD several a of form composed that typically is fold repeat tandem WD40 The contain Mps1. multiprotein and repeat that BubR1 WD40 of Bub1, are proteins assembly motif this checkpoint the of arrangements a for Mitotic a as functions module from generally complexes. it ten, proteins, organizing than more TPR-containing versatile to repeats TPR between two of of number greatly tandem the Although varies plants functions. and of units animals diversity from a proteins mediate in of that distributed packing widely the is from helix-loop- motif results TPR a that of in twist structure super-helical organized entire right-handed the are a define that repeats motif acids TPR a helix. amino defined 34 motif protein of a is consensus motif (TPR) repeat tetratricopeptide The motif TPR are Knl1 that with domains interact protein to of known features General 2. Box nadto ovraini ubr eunedvriyo the of diversity sequence number, in variation to addition In b poelrbigtems omntp.Teblades The type. common most the being -propeller hs eet r paetydsesbefor dispensable apparently are repeats these , b srn foerpa neat ihthe with interacts repeat one of -strand b seto ld.Ec WD Each blade. or -sheet , 0aioacid amino 40 a hlcs The -helices. D. oelgtot h oeua neatosudrinn SAC underpinning interactions molecular shed will the the regulation. recruitment onto how BUB light of coordinates more studies motif of structural MELT characterization The and extended important biochemical the are tolerated. motif and this and regulates discoveries, be that motif kinase principal the MELT might as an Mps1 the that groups suggesting of phosphate complexity, identification structure of low of incorporation be to predicted einecmasn h T the Mps1 encompassing Knl1 the and/or addition, In region Plk1 modules. MELT a most in resembles site phosphorylation loosely sequence Y)-(ST)-(DE) ieohrsi re obn oKl.I gemn ihthis, with Bub1 agreement on In dependent strictly Knl1. of is to context to bind al., the recruitment to et BubR1 requires (Zhang order BubR1 direct in is Bub1, kinetochores interaction unlike this whether Moreover, clear 2014). peptide, not is phosphorylated-MELT-containing it a and but Bub3–BubR1 with Importantly, Bub1 associate remain. of can still recruitment questions the many for BubR1, motifs MELT phosphorylated motif. of affinity individual the MELT the by to of chromosome qualitatively Bub3 number and phosphorylated given the the permutations are any by that motifs way, quantitatively at and determined sites this are BUB-binding response moment In functional of SAC needed. combinations and as the machinery fine-tune the MELT congression allows that of to al., ‘rheostat’ sequence a as et and cell serve number might (Vleugel sequences exact these they clarification the motifs, promote how of further to Regardless and awaits 2013). sequences species, recruitment all MELT in BUB phosphorylated conserved with not two However, coordinate 2013). are the al., et features (Vleugel Interestingly, Bub1 cells these of human in end. recruitment kinetochore for the essential C-terminal to be to the appear sequences at flanking SHT sequence T sequence where the by side (ST)-(DE), N-terminal the on flanked often erimn?I ua n1 h henn eiu fteT BUB the of on residue a effect threonine the an Knl1, there T have human In example, this recruitment? that does for is the and motif, motifs Are phosphorylated MELT or regulation? (ST)-(DE), MELT the Mps1 of Mps1, different of sequences layer flanking for for additional an sequences Mps1 substrates constitute could of MELT as affinity all suited differential and are functionality well SAC However, equally for substrate alignment. Mps1 crucial chromosome has a motifs MELT MT-binding as the its of emerged residue regulate phosphorylate threonine thus the Bub1 addition, and In domain activity? Can MT-binding Knl1. its at and Knl1 Bub1 feedback a between is there mechanism whether of its question the understand raises this biorientation; fully to order docking. in kinetochore Bub1, of through with achieved mechanism is is interaction BubR1 this how direct whether precisely and a answering kinetochore the so, on Future to focus if regulated. recruited to and, is need stochiometry proteins MELT will checkpoint 1:1 of these efforts a BubR1 of occupancy Chen, obey binding the BubR1 the of 2002; how and whether al., independent Bub1 is by et is question motifs Millband related Bub1 A 2002; of 2002). Hardwick, that and whereas (Millband Bub3, and eiuso h eunepeeti ua n1 r oae near located are two Knl1, first human consensus in the present the sequence after the by named of residues defined but motifs, KI(D/N)FxxF(L/I)xRL sequence (lysine-isoleucine) KI Two motifs KI lhuhteercn tde l eonz h infcneof significance the recognize all studies recent these Although oevr u1kns ciiyi eurdfrchromosome for required is activity kinase Bub1 Moreover, ora fCl cec 21)17 4532 doi:10.1242/jcs.149922 3415–3423 127, (2014) Science Cell of Journal W eoe yrpoi eiu,adthe and residue, hydrophobic a denotes W (/)(T-D)sqec is sequence -(F/Y)-(ST)-(DE) WW W WW -(F/Y)- -(F/Y)- -(F/

Journal of Cell Science ihaprxsmltreigsga- PS)cgaepeptide cognate (PTS1) signal-1 targeting complex peroxisomal in PEX5 a and in respectively); with STIP1) 1ELR, and peptides as mimic 1ELW HSP90 IDs known and (HSC70) (PDB 70 also shock heat 2BUG); (HOP, a ID with protein complex (PDB organizing (HSP90) in 90 shock (PP5) protein shock heat 5 heat phosphatase a protein several with in complex as observed [such binding ligand complexes of TPR–ligand mode TPR the the the of to arrangement that conforms tandem the motif considering by defined Indeed, is and that face partners. and motifs concave KI interchangeable binding respective not other their are for potentially the specificity BubR1 that evolved suggesting and likely another, Bub1 have for of substitute domains can KI1 TPR motif the neither KI2 Remarkably, the 2011). nor al., et (Bolanos-Garcia Knl1 (I nefc htivle eiusI hydrophobic residues complementary involves extensive that BubR1 similar of interface a domain TPR shows the KI2 of interaction with The 2012). al., et (Krenn COMMENTARY hsdfnn P oan(i.2,) ntecs fBub1, of case the (I In 2C,D). residues (Fig. tandem domain a 2), TPR conserved (Box by and a motif mediated Bub1 defining (TPR) is repeat of thus tetratricopeptide respectively, binding the KI2, The of and 2011). arrangement KI1 al., (Kiyomitsu et to Bub3–BubR1 Bub3–Bub1 Kiyomitsu BubR1 of the 2007; that of al., (KI2) recruitment et second the features; the motif functional mediates and unique MELT (KI1) complex shows first one motif KI first the Each the of 3). downstream Fig. 1; Knl1, (Fig. of N-terminus the C A Bub3 TPR-Bub1 177 KI2 xTxxFLxxL KI1 – GLEBS-Bub1 Bub3 – 186 GLEBS-Bub1–Mad3 eiea xesv yrpoi interface hydrophobic extensive an define ) TPR-BubR1 – pMELT 177 a potentialadditional TPR concaveface: ,T ligand-binding 179 region 213 ,F ,F 215 182 ,F B Bub3 n L and D 218 ,I – GLEBS-Bub1 219 F175 186 n L and )ofKI M171 T172 222 M169 Nsl1 of p – pMELT – Knl1 P166 T167 eetees lhuhi per htteK oisare motifs KI the that 2012). promotes al., appears that et se it (Yamagishi robust per although the BubR1 KI1 for of necessary Nevertheless, with not recruitment interaction is KI2 kinetochore the suggesting Similarly, 2012), not activity. al., catalytic KI1 is et Yamagishi with it 2012; interact al., that to et localization al., (Krenn unable kinetochore et Bub1 or for is activity of Ricke kinase that necessary altered 2012; mutant exhibit be not al., TPR does to et Bub1 reported (Krenn a activity been 2012), an kinase has is although Bub1 domain Moreover, Knl1 optimal 2012). TPR al., in Bub1 et KI2 Yamagishi the In alignment 2012; nor chromosome al., 2012). or et KI1 al., activity (Krenn of SAC et for presence to requirement Yamagishi Bub1 absolute the of 2012; neither recruitment al., the TPR addition, et for the sufficient (Krenn with not the KI1 kinetochores is of of Bub1 context interaction of the in the domain However, molecule, 2013). al., Knl1 al., et (Vleugel et full-length recently Vleugel Vleugel evolved 2012; 2012; has al., motif al., et KI et the that Krenn 2011; suggesting 2011; al., 2012), et al., (Kiyomitsu et orthologs Bolanos-Garcia Knl1 vertebrate in confidence through partners binding additional 2C). (Fig. recruit domains interfaces TPR BubR1 alternative the that and possible is Bub1 it 2012), of (Bolanos-Garcia al., et fold Krenn TPR 2011; al., the et the of at principally interface occurs BubR1 convex and non-canonical Bub1 2011)], to binding Blundell, Knl1 and that Bolanos-Garcia and in (revised 1FCH) ID (PDB h rsneo I n I a nybe dniidwith identified been only has KI2 and KI1 of presence The ora fCl cec 21)17 4532 doi:10.1242/jcs.149922 3415–3423 127, (2014) Science Cell of Journal eitsKl idn oNl PBI 4NF9). ID and (PDB motif Nsl1 RWD to is the binding Knl1 of Knl1 of arrangement mediates region tandem C-terminal double The as (D) organized conformational binding. ID little (PDB Knl1 undergo motif upon KI2 TPRs changes Knl1 both ID the that (PDB and showing motif BubR1 KI1 3SI5), from Knl1 TPR the the and and Bub1 4A1G), from binary TPR the the of of the complexes superposition on Structure ‘roof’ (C) a MELT. define phosphorylated residues and N-terminal interaction residues the Bub1 few to that that contribute motif shows MELT complex phosphorylated ternary the state. of the ordered of an structure to The important disordered (B) undergo a to from likely changes is conformational peptide complex mimic upon pMELT contrast, the In formation of motif. binding MELT upon phosphorylated Bub3 that in the occur showing changes 4BL0), conformational ID minor (PDB only of motif) mimic MELT peptide a phosphorylated is the pMELT Bub3–Bub1-GLEBS– (where the complex ternary and motif 2I3S) pMELT ID motif (PDB Bub3–Bub1-GLEBS complex the of binary superposition Structure Bub3. in complex kinetochore and SAC components. of Structures 2. Fig. A w rti-idn oishv enidentified been have motifs protein-binding Two (A) 3419

Journal of Cell Science COMMENTARY nierdfamn,wt h aaiiyo nacdBub1 enhanced of 3420 capability the with non-functional KI2 otherwise fragment, this and provides from KI1 Knl1 engineered repeat of MELT of in region functional central inclusion is single the a Strikingly, which of a 2013). downstream manner, sequences al., loading cooperative to kinetochore et a for Bub1 dependency (Primorac MELT1–KI in mutual and their occurs phosphorylated with agreement Bub3 Knl1 the of of 2012). mimics region al., binding that et concerted (Krenn phosphopeptide interactions the Bub1–KI1 Moreover, Bub1 stabilize BubR1 reduced the between KI2 in interactions in that and results motif idea fragment the KI2 Knl1 prompting BubR1-specific recruitment, N-terminal a the study of kinetochore of one context Interestingly, removal the 2013). that al., the to et reported promote Vleugel proteins 2014; to al., checkpoint et repeat (Krenn mitotic MELT cooperate single of motifs adjacent KI recruitment both the that MELT shown with have first strongly KI2 the and only KI1 and encompass motif that 2013). fragments al., et Knl1 (Vleugel N-terminal cases both of in amounts when kinetochore similar the to recruiting to compared despite Bub1 present, alignment is metaphase 5- Knl1 a in 150– full-length in residues delay al., results Knl1 MELT2) 10-minute of et and of to K2 deletion Vleugel efficiency KI1, notion, MELT1, the 2014; (encompassing this the 300 of al., at to support et proteins In contribute 2013). (Krenn BUB still congression of might chromosome presence they overall kinetochore, the for 2014). dispensable Biggins, and (London Mps1 by the Hs, protein involves organisms. latter kinetochore diverse the the from of to respectively, phosphorylation recruitment motifs, the Mad1 KI2 requires Mad yeast, and process the as in KI1 the and/or MELT, such that, that the BubR1 components, shown and of and SAC glaber has Bub1 Bub1 alignments other Hetecephalus laboratory of between sequences Furthermore, Biggins region acid interaction investigations. the middle Bub1, Amino direct further from catalytic the with (B,C) the require work the to interactions mediate that example, stimulate Mad1 The and aspects For can of motifs. kinetochore, important involved. KNL1 binding KI the are be cartoon in the to sites, also the rearrangements of BubR1 KNL1 simplicity, might structural downstream of additional For Mad2, such located recruitment to Bub3. Whether are the Bub3 and KNL1. that to of BubR1 in KNL1 contribute binding Bub1, changes human Bub1, to conformational in of binding local repeats domain sustain MELT important kinase and putative induce promote few Bub3 to a and motifs only BubR1 MELT of Msp1-phosphorylated phosphorylation with the cooperate shows can KNL1 repeats. of motif MELT motifs of Function 3. Fig. h euneainet niaeta hs ucinlmtf r ihycnevdi hs organisms. these in conserved highly are motifs functional these that indicate alignments sequence The tde fKl rnain aecm osmlrconclusions: similar to come have truncations Knl1 of Studies A B As Xt Bm Ch Cl Pa Pt Hg Mm Hs MELT Bub3 Mad1

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trpsalecto Pteropus T MELT motifs V V V V V V I A V V ? L MI M VI I I I I I I I P T T T T M T T A T K K K K K K K K K P Cl, ; C P ai lupus Canis Xt Bm Ch Cl Pa Pt Hg Mm Hs As 281 L I K I K K K TP ET K K Knl1/Spc105/CASC5 S S S E N S S N S E E G E E E E E E D Bm, ; K K KS K KS N Q K K K S S ST S S S I S S P oan swl steBb-idn ein ilg long a go will issue. region, this Bub3-binding solving in the to the as complex way both well includes as KI2 that fragment domain, Bub1 and TPR a 2004; and KI1 Bub3 al., the with of to MELT, association et studies contribute structural phosphorylated (Johnson also future Undoubtedly, N-terminal might kinetochore 2009). Knl1 al., the for the et in Klebig of to motif KI1 Recruitment recruitment to MELT Bub3. Bub1 BubR1 adjacent to of whether binding the domain and investigate and/or TPR primes Mps1 presenting to kinases Knl1 by important in specific recognition to be important to binding will Knl1 Bub1 be it of and might regions phosphatases, transition flexible al., a et unbound (Bolanos-Garcia Such binding BubR1 2011). upon Knl1 a peptide domains of and binding, mimic transition Knl1 TPR disorder-to-order KI2 Bub1–Knl1 a upon reported two change have the we conformational the although little of a that only structures indicate undergo the complexes with BubR1–Knl1 domains apparent, readily studies. BubR1 not structural from is emerged achieved have binding. is clues of this enhancers some as exactly although act how motifs KI2 of and Details KI1 support the collectively that results view These the function. SAC and recruitment faioai eiu osrain hsrgo sognzda a as organized is region degree This high conservation. a residue shows acid Knl1 amino of of region C-terminal the metazoans, In domain RWD

T S I T T T T N T R o mutus Bos uepsto fte3 tutrso P u1adTPR and Bub1 TPR of structures 3D the of Superposition K K K K K K K K K K I I I I I I I I I I D D D D D D D D D D ora fCl cec 21)17 4532 doi:10.1242/jcs.149922 3415–3423 127, (2014) Science Cell of Journal

T F T T T V T T T 290 T A T T T T T T T T KI1 Ch, ; S S S S S S T S S F F F F F T F F F F F M L L ar hircus Capra L L L L L L L N A A A A A A A A A E S N N N N N N N E N L L L L L L L L L L K K K K K K K K K A H R L L L L L S S L 299 Xt, ; eou tropicalis Xenopus oosapiens Homo E N K K K N K 334 K T K K K K K K K K K K K I I I I I I I I I I N N N H N N N N D D F F F S S F F F F F K N N N N N N N N D E D D D D N D D KI2 D D 340 ;Mm, F F F F F F F F F F I L I I I I L I I As, ; I K K K K K T K M K K u musculus Mus G R R S R R R R R R liao sinensis Alligator L V L L L L L L L L K K K K K K K K K K T I S I T T T T T T G G G G E G G G S G K K K K K K K K K K 349 and 1 Hg, ; .

Journal of Cell Science YxP hti eie ytersde Y residues the by defined is motif that YPxxxP non-canonical a contains (YPxxP) RWD Interestingly, the the 2012). as Knl1, 4NF9, al., human well et in as (Kim 2014), and Mad1 al., protein that et checkpoint 4NFA Petrovic mitotic RWD-fold 2013; al., (PDB a et and Ctf19 (Nishino adopts Spc25, Mcm21 Nsl1 Spc24, Csm1, region to features with structural the similar shares that complex confirm in The respectively) 2012). domain and C-terminal Harrison, Knl1 the alone and of structures Schmitzberger of crystal stability reported 2004; protein Mutation recently impairs al., 2012). often et Harrison, motif (Nameki (Nameki YPxxxP and few the Schmitzberger within a residues name 2004; (also to al., GCN2 Spc25, of et and domains Spc24 RWD proteins the the kinetochore and (E2s) of enzymes ubiquitin-conjugating structures EIF2AK4), as 3D known the (triple in residues two seen by other each with overlap consecutive that three turns includes that loop stable a forms motif YPxxxP oe.Freape motn oa ofrainlcagsin Y be the residue changes surrounding can conformational region is differences RWD local that Knl1 important complexes important the time, interactions example, these same For the hydrophobic noted. of At cooperative them. analysis stabilize the feature of common from Another predominance 2014). emerges formation al., upon et turn that (Petrovic helical the tight complex In a the of 2014). of transition formation al., disorder-to-order the et a in undergo results Petrovic that to 2013; appears al., Nsl1 case, et latter Primorac 2012; al., et n1K oi rBBadteML oi nysmall only the motif of MELT formation the after and observed BUB the are or and changes BUB motif between conformational complexes KI the in Knl1 whereas binding, Nls1 eua 03.TeRDdmi,wihi ae fe three after named is which domain, and and (Caldas RWD complex assembly The Mis12 kinetochore 2013). the proper the Both DeLuca, for mediates 1). required domain (Fig. (Petrovic are coil Zwint Zwint Nsl1 to coiled physically Knl1 Knl1 component domain of the binding complex RWD whereas 2014), The Mis12 al., 2D). 2010; et the Fig. al., with et 2014; (Petrovic interacts region al., coil et coiled of a Petrovic presence and the domain by RWD characterized the is that domain globular compact COMMENTARY eety h n1RDdmi ncmlxwt l1mimic more Nls1 and a motifs with complex MELT Q in Knl1 (residues domain KI peptide to RWD the Knl1 bound with complex the Bub3 in recently, BubR1 Knl1, and of Bub1 of motifs domains that including TPR complexes, the diverse of of structures the from emerging KMN the and co-evolved. SAC have the network of modular components that central speculate of regulate to motifs functional tempting that is Mps1 it and segregation, BubR1 chromosome recurrent the Bub1, of kinases feature a common that checkpoint a is mitotic Considering are motifs motif architecture. repeat of kinetochore arrangements RWD tandem of the module that structural suggesting species, and RWD across triple of the architecture including the motifs, structural E2s to this overall binding whether The its define complex. including domain to Mis12 function(s), important Knl1 RWD to be the contributes would motif of it binding, rearrangements Nsl1 (by local upon region important Nsl1-binding the located the is given motif from YPxxP away a Such far S1). Fig. material (supplementary (residues motif YPxxxP the of presence the L is DEAD domains yeast RWD as an finger-containing organized with and typically fold RING is proteins helicases, (DEXD)-like proteins: WD-repeat-containing proteins, RWD-containing major 231 atr fdsre-oodrtastosi A inln is signaling SAC in transitions disorder-to-order of pattern A PSPYP 236 a - nCf from Ctf9 in b - b 266 - b - –P b - a 274 - a BlnsGri ta. 01 Krenn 2011; al., et (Bolanos-Garcia ) oooy n tutrlfaueof feature structural One topology. lyeoye lactis Kluyveromyces b tr,i udmnal conserved fundamentally is -turn, 2125 , 9A 19 aepaeupon place take a ˚ .Otn the Often, ). 2262 + .However, ). b b PSVP sandwich tr) as -turn), 2266 b - emnlrgo fKl n o uhcnomtoa changes conformational signaling. SAC such modulate how to is C- and contribute the Knl1 it by can mediated of Similarly, interactions region the domain. underlie terminal domain rearrangements RWD RWD structure the local the of important other similar by RWD-containing whether extent mediated unclear involve what that to are network establish proteins KMN to the important of be interactions will It complex. eit rdcieitroeua neatosi crucial a that is surfaces flexible interactions highly intermolecular signaling. and SAC productive large on has of mediate consequences expression establishment the Knl1 detail of an The it greater suppression 2000), of in the al., function study that et to the Albert interesting on 2005; be of effect would (Albert, stability profound network the a interaction of have entire impairment can that or proteins surprising expression not hub is the it Although of 2006). Kim al., suppression 2012; et Haynes al., 2008; et al., proteins et (Babu Doszta hub of networks 2006; of al., interactome frequency feature et in a high found is A are complexity that structure chain. low polypeptide distributed of shows are regions entire Knl1 that domain, complexity its RWD structure throughout low globular of the regions of multiple exception the With for implications – signaling Knl1 cell of complexity structural Low 02 oe n aor 03.I slkl htatvto and activation to that response likely in kinetochores is at It proteins checkpoint 2013). al., of Kapoor, inactivation et and (Lara-Gonzalez Foley kinetochores unattached 2012; levels at maximal reached with microtubules, being to kinetochores the of during of regulated attachment levels dynamically are The kinetochores SAC. at the proteins in and SAC role to KMN important the tempting an between have communication is interactions the It of types 2012). these al., that speculate al., et underlie et (Pan that Miermont processes events including 2010; mechanotransduction 2005; 2005), Elcock, Wright, Fradin, and and and Dyson McGuffee 2013; (Banks 2012; remodeling al., and et Cino stability their cell as as complexes, the protein well of of association environment of an kinetics crowded the influence the might mount in complexity to structure low framework structural a response. SAC provide effective of defined a mode should to cooperative concentration region possibly local and al., and colocalization underlie et coordinated protein Rosenberg interaction, a that 2010; such interactions al., In to N-terminal et appears 2011). concerted the KI2 (Liu of and to transitions KI1 mapped disorder-to-order mode to proximity been a close have in support for are sites that Knl1 recognition B of the region that might Aurora fact The and BubR1 important SAC. is PP1 the or that in function process Bub1 to its a Knl1 for phosphatases, of of regions and/or unbound kinases binding and specific flexible of upon exposure the Knl1 facilitate of mechanism sequential transition or concerted their a vivo to in follows BubR1 and motifs Bub1 their KI of enhancing binding thereby specific the BubR1, Whether of Bub1, Knl1. domain of to TPR domain binding the TPR to the to KI2 for motif of KI case and Knl1 the to the contribute be of could presentation well which the might Mps1, This by of phosphorylation 2013). macromolecular al., motif structure al., MELT of et low et number regulation Nishi of (Xue and is regions pathway assemblies formation of a Wnt role the the for the of of complexity (Peng defining review excellent those proteins assembly an and 2012; ribosome 2014) proper as al., et such the complexes, macromolecular for requirement h soito fpoen hog h neato frgosof regions of interaction the through proteins of association The ean ob salse.Mroe,tedisorder-to-order the Moreover, established. be to remains ora fCl cec 21)17 4532 doi:10.1242/jcs.149922 3415–3423 127, (2014) Science Cell of Journal ´ y ta. 06 ukre l,20;Dunker 2005; al., et Dunker 2006; al., et nyi 3421

Journal of Cell Science akta ilrqieitnie–adide netv research – mitotic inventive indeed in and – challenging functions intensive efforts. molecular and require important will Knl1 that an the signaling. task remains of SAC thus signaling into of understanding time checkpoint events unattached molecular and early insights the space to The regulate in fundamental that response organized mechanism(s) provide are in these the triggered should how precisely Knl1 and interactions of more kinetochores understanding for Knl1 An account define organisms. the different that to in motifs recognition important partner species-specific be of will requirement the across above, It considerably discussed varies multiple as modules species. those although, of protein, of sequence and this number to involvement unique greater are the that as by segregation, gained chromosome be of components. control can essential have the selectivity might in components in complexity KMN structure roles and low proteins of SAC regions and other Furthermore, productive environment cell. numerous, crowded the the the in of be undergoes In of to it that likely response. establishment interactions is specific RWD the Knl1 cellular Knl1 of for the the flexibility by crucial structural mediated of intrinsic are the that regulation domain, interactions the of thus to ligands, layer addition more or a one with providing interaction the mediate that motifs CDKN1B CDKN1A ltomta speetdb n1mgtidc allosteric induce further deserves might that possibility the investigation. interesting Knl1 at an activities is by This multiple docking kinetochore. coordinate the presented that to changes is conformational proteins the that SAC for cellular of platform dynamic crucial binding highly be and Furthermore, intricate to process. essential, prove this of will regulation events dephosphorylation phosphorylation and through attachment kinetochore–MT of status the COMMENTARY 3422 V. R. Pappu, N., and W. Grigorieff, R. A., Kriwacki, D. M., Ball, M. S., Babu, Pasqualato, H., V. Ramey, M., G. L. Alushin, A. Barabasi, and H. Jeong, R., Albert, R. Albert, References at http://jcs.biologists.org/lookup/suppl/doi:10.1242/jcs.149922/-/DC1 online available material Supplementary material award. salary Supplementary Investigator holds New S.E. Sciences Health Research. by of and supported Health Institutes is of Canadian S.E. Institutes a Canadian of the Groome laboratory from (Nigel the funds support in operating financial Work the Studenship). for Research University University Brookes Oxford thanks M.K. Funding interests. competing no declare authors The interests suggestions. Competing and comments insightful their for referees anonymous the thank We Acknowledgements p21 are interactions of Examples classes regulation. cycle these cell to in interactions of response specific, effective highly an but mediate affinity, low binding through with partners associate frequently proteins disordered Intrinsically perspectives and Conclusions estlt rmpoendisorder. protein from Versatility microtubules. along arrays oligomeric E. Nogales, and A. Musacchio, networks. complex otKl oooscnana ra frpaigmodules repeating of array an contain homologs Knl1 Most 20) cl-rentok ncl biology. cell in networks Scale-free (2005). oCk–ylnA hsotnivle utpelinear multiple involves often This Cdk2–cyclin-A. to ) idn ocsi iaeI,adp27 and II, kinase casein to binding ) Nature 406 378-382. , Science 21) h d8 ieohr ope forms complex kinetochore Ndc80 The (2010). Nature 337 20) ro n taktlrneof tolerance attack and Error (2000). 1460-1461. , 467 805-810. , .Cl Sci. Cell J. 21) tutrlbiology. Structural (2012). WAF1 KIP1 ecddby (encoded 118 ecddby (encoded 4947-4957. , oao-aca .M,Lshti . Matak-Vinkovic T., Lischetti, M., V. Bolanos-Garcia, C. Bolanos-Garcia,V.M.andBlundell,T.L. Fradin, and S. D. Banks, ans . lfed .J,J,F,Kigr,N,Csc,M . aioa,P., Radivojac, E., M. Cusick, N., Klitgord, F., Ji, J., C. Oldfield, C., Haynes, Sillje M. A., Hanisch, Gstaiger, and R. Aebersold, A., Wepf, T., Glatter, i,P . u .J,Xa .adGrti,M B. M. Gerstein, and Y. Xia, J., L. Lu, M., S. P. S. Taylor, Kim, and D. Hussein, V., S. Holt, F., I. M. Scott, L., V. Johnson, ads .V,DLc,K .adDLc,J G. J. DeLuca, and F. K. DeLuca, V., G. Caldas, G. J. DeLuca, and V. G. Caldas, ei,A,Dsr . abr,N,Bevit . a e a . irun G., Pierquin, N., Aa, Der Van M., Biervliet, N., Lambert, J., Desir, A., Genin, W., E. P. J. Wright, Chen, and Y., J. J. H. Dyson, Yang, P., Romero, J., Meng, J., Uversky, C. and Oldfield, M. K., A. L. Dunker, Iakoucheva, P., Romero, S., M. Cortese, K., A. Dunker, Doszta Mu S., A. Rybina, Musacchio, A., Desai, and G. J. DeLuca, am .R,We,J . ail .J,Svkmr . cvy .N,Potapova, N., J. McAvoy, S., Sivakumar, J., J. Daniel, D., J. Wren, R., J. Daum, W.-Y. Choy, and M. Karttunen, A., E. Cino, A. Desai, and T. Fukagawa, T., Hori, M., I. Cheeseman, A. Desai, and M. E. Wilson-Kubalek, S., J. Chappie, M., I. III, Cheeseman, R., J. Yates, F., Hyndman, S., Anderson, S., Niessen, M., I. Cheeseman, A. Santamaria, and A. E. Nigg, A., A. Jeyaprakash, W., Y. Chan, atns .N,Snaai,A,Jypaah .A,Wn,B,Cni .and E. Conti, B., Wang, A., A. Jeyaprakash, A., Santamaria, N., T. Gaitanos, J. D. Wynne, M. and H. Funabiki, T. Kapoor, and M. Maldonado, A., E. Foley, hn .H. R. Chen, set . heabtu,D . rnig . eea .adDsi A. Desai, M. T. Kapoor, and and A. E. K. Foley, Oegema, L., Krenning, K., D. Cheerambathur, J., Espeut, utfctdknsso h elcycle. cell the of kinases multifaceted crowding. molecular vrk,V . ia,M n aocea .M. L. Iakoucheva, and M. Vidal, N., Ska2. V. Uversky, and Ska1 comprising complex kinetochore and J. spindle novel a 5306-5317. iesoa tutrst rti ewrspoie vltoayinsights. evolutionary provides networks protein to Science structures dimensional interactomes. eukaryotic four from proteins hub Biol. of Comput. feature common Chromosoma Site. binding unanticipated an via Structure regulation checkpoint kinetochore-mitotic Blundell, for and J. Nilsson, V., L. C. Robinson, T. R., D. Spring, Y., D. Chirgadze, J., P. oklwfrcatn h ua neato rtoe nihsit h PP2A the into insights proteome: interaction human the system. charting for workflow al. et microcephaly. A. primary Lefort, in mutated M., CASC5 Urbina, M., network Tosi, A., Killian, proteins. disordered intrinsically N. on V. Uversky, update and an Z. Obradovic, V., Vacic, network for implications networks. their N. and V. proteins hub in repeats evolution. sequence and in kinetochore the of interface binding K. Oegema, and A. Hyman, 19 u1i eurdfrkntcoelclzto fBb1 epE epFand Cenp-F Cenp-E, BubR1, of congression. localization chromosome kinetochore and for Mad2, required is Bub1 .A n obk,G J. G. Gorbsky, and A. T. proteins. disordered intrinsically of dynamics the on 487-496. tension. sustain to ability its and kinetochore 18 A. outer the Desai, of assembly and K. Oegema, complex-KMN Ska interaction. inhibiting network by attachments kinase kinetochore-microtubule B controls B Aurora promoting by proteins kinetochore activity. outer of phosphorylation iecn n h aneac fcrmsm oeini mitosis. in cohesion chromosome of maintenance the and silencing k ope n t e opnn Ska3/C13Orf3. component new its and complex Ska A. E. Nigg, B56- dephosphorylation. the and phosphorylation by on depends microtubules and phosphatase. 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