Journal of Cell Science Australia eet nnuoa rgntrcl iiinaergre as regarded are 2009). Woods, division and cell (Thornton which development progenitor disease in neuronal underlying with (MCPH) in mutations microcephaly protein defects primary centrosomal recessive linking with autosomal Proteins studies 2009). revealed genetic been Gergely, have by regulation and centrosome/spindle Zyss in roles 2009; diseasecritical human Raff, to and linked spindle are (Nigg and bipolar progression cycle aberrant cell in formation, in defects Importantly, result 2011). al., functions et centrosome-directed of Megraw 2010; plane al., cells the daughter et into (Lesage specify segregation cellular to precise dictate proper required and division and are activity centrosomes astral MTOC of Moreover, positioning 2004). facilitates al., al., et et that (Fong Zimmerman Lu 2009; assembly al., spindle et cdk5rap2) Haren bipolar 2008; and and nucleation the microtubule via pericentrin PCM is the NEDD1, centrosomes of at duplicated of expansion early considerable of anchored recruitment the separation a anti-parallel In by and the accompanied (PCM). mitosis, in poles of material stages pericentriolar microtubules the proteinaceous surrounded spindle at centrioles by paired of focused of consist Centrosomes centrosomes. are ends arrangements minus microtubule-based mitosis, During organized the 2000). (Compton, division chromatid highly cell sister during faithful a segregation in functions is essential with superstructure spindle bipolar The Introduction neurogenesis. coordinating in involved be words: may Key that regulation spindle WDR62 mitotic of in characterization functional signaling first JNK/WDR62 the for requirements provides revealed is study WDR62 has Our of mitosis. and phosphorylation during of JNK organization revealed integrity spindle we the metaphase Additionally, maintaining progression. decreased in mitotic defects, for function delayed orientation required and spindle WDR62 pole revealed in spindle resulted we the loss from depletion, WDR62 displaced centrosomes siRNA metaphase. Utilizing during specifically transition. metaphase–anaphase spindle mitotic spindle until the the persisted stabilizing at that WDR62 entry of 62 accumulation mitosis cell-cycle-dependent mitotic demonstrated protein in we during roles WD40-repeat cells, pole cultured its determination. In but capacity. the microcephaly fate proliferative that their of reduced cell here defect neurodevelopmental and report and exit the We progression to defined. linked mitotic protein been pole for not spindle have required a of regulation as is tight identified the recently that indicates was development positioning (WDR62) embryonic and in division formation cell asymmetric spindle on bipolar spindles and/or centrosomes aberrant of impact The Summary 10.1242/jcs.107326 doi: 5096–5109 125, ß Science Cell of Journal 2012 July 4 Accepted ( correspondence for *Author 2 1 Bogoyevitch A. Marie progression maintenance mitotic spindle timely for and required protein JNK-phosphorylated pole a spindle is 62 protein WD40-repeat 5096 uinI Heng I. Julian eateto iceityadMlclrBooy i2 oeua cec n itcnlg nttt,Uiest fMlore itra3010, Australia Victoria 3010, Melbourne, Victoria of University, University Monash Institute, Institute, Biotechnology Medicine and Regenerative Science Australian Molecular The Bio21 Biology, Molecular and Biochemistry of Department 02 ulse yTeCmayo ilgssLtd Biologists of Company The by Published 2012. etooe pnl oe Drpa rtis ioi,Cl iiin Neurogenesis division, Cell Mitosis, proteins, WD-repeat pole, Spindle Centrosome, c tbln and -tubulin- 2 n oii .H Ng H. C. Dominic and [email protected] dr ta. 06 sioie l,2009; al., et Oshimori 2006; al., et ¨ders 1 vneY .Yeap C. Y. 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WDR62 (Fig. cytosolic interphase HeLa during distinct a WDR62 in recognizing revealed analysis antibodies consistently Immunofluorescence using analyzed cultures. we cells immunofluorescence, cell by not human protein but human WDR62 detected murine As antibodies intracellular cycle. available the cell commercially WDR62 of the three analysis throughout our WDR62 In endogenous prompted centrosome of of 2010) distribution proteins. al., shared et localization Yu MCPH 2010; the multi-compartment different with (Bilgu the the consistent for regulatory addition, cycle observed is cell association and important suggests functions capacity neuroprogenitors proliferative the sustaining of in WDR62 for requirement A spindle protein a pole as localization WDR62 Cell-cycle-dependent eeucagd(i.3) n elto a utie o at S2A). Fig. for material sustained (supplementary was post-transfection depletion h and 72 of 3A), least (Fig. levels a unchanged to whereas compared were siRNA when as Con individual WDR62 Four division, silenced non-targeting siRNA. substantially cell with mitosis, cells all during in HeLa siRNAs regulation in WDR62 temporal WDR62 of and depleted we spatial roles its potential by suggested the define mitosis of To completion delays depletion WDR62 aeilFg 1) ncnrs,w bevdpoietWDR62 prominent with observed we contrast, colocalization In S1A). Fig. material ooaie ihtemcouuecoslne,ncermitotic subunit nuclear component, WDR62 complex p150 dynactin cross-linker, regulatory the contrast, and microtubule protein, In (NUMA) apparatus cdk5 the 2D). 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(Fig. poles spindle at , tblnsandcnrsmsbgniga prophase at beginning centrosomes -tubulin-stained D6 ucini ioi 5097 mitosis in function WDR62 c tblni ioi el indicating cells mitotic in -tubulin c tblnc-tiig(supplementary co-staining -tubulin a and - c -tubulin Journal of Cell Science BrdU rlfrtn el (GFP cells proliferating GFP GFP h rprino el obelbldfrGPadpH (GFP pHH3 and GFP for labeled double cells of proportion the rae mro.( embryos. treated eemndb onigGFP counting by determined t,t,t n 4,acmie os D6 iN ol o-agtn iN (C siRNA non-targeting pool, siRNA WDR62 mouse combined cortex. a embryonic t4), the and within t3 proliferation t2, cell (t1, decreased depletion WDR62 1. Fig. 5098 lcrprtdwt oto rmueWR2sRAwr tie o hsh-itn 3(H3.Mgiidpnl blw fteV ihih el nMp M in cells highlight VZ the of (below) panels Magnified (pHH3). 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HeLa of duration in the altered not division depletion prevent WDR62 mitosis. that or were found we arrest contrast, not cycle did viability levels loss cell WDR62 WDR62 induce Thus, phenotype. cell S2B,C). reduced Fig. substantially knockdown material (supplementary despite and the 3 altered of target markedly proliferation other specificity and two 2 ensure Interestingly, least target at to WDR62 with siRNA) (typically re-confirmed were WDR62 siRNAs but by independent depletion siRNA WDR62 1 utilized target findings subsequent All hnteefc fWR2ls a vlae nsynchronized in evaluated was loss WDR62 of effect the When Fg D rknwielns.Rfetn hsmttcdly a delay, cells mitotic WDR62-siRNA-treated in this index of Reflecting mitotic in suggestive lines). mitosis increase prolonged white observed modest of broken was periods 3D, during plate (Fig. defects metaphase orientation the spindle 3E) and of (Fig. movement substantial mitosis rotation Furthermore, complete arrest. WDR62- to cycle cells) cell without Some control of took 3E). duration Fig. average cells with min; compared (54 when with nearly depleted mitosis cells 3D,E) complete average (Fig. to min) on Con-siRNA-treated prolonged (96 taking long was as cells twice post- mitosis showed HeLa h of 4 further WDR62-siRNA-treated duration by cells nocodazole the WDR62-siRNA-treated markers Time- that from of mitotic 3C). 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Fig. a:20 bar: Scale NPC). (Mab414 proteins complex nuclear pore with co-staining by as demonstrated breakdown envelope nuclear prior to occurred centrosomes with association 20 bar: Scale poles. spindle and centrosomes revealed immunostaining E D6 olclzto ihspindle with co-localization WDR62 ) m .( m. C ihrmagnification Higher ) D osann of Co-staining ) c m -tubulin m m .Ist are Insets m. .( m. m. ( Glued A B WDR62 ) WDR62 ) .Insets ). c - Journal of Cell Science a oe by microtubules noted WDR62 mitotic and astral prophase (as of anchoring in centrosomes result assembly delayed intact spindle a the of prometaphase, as examination examined underlying closer organization On we spindle loss. defects, mitotic features in orientation changes spindle the and progression interrogate a knockdown indicating WDR62 To with mitosis defects spindle of Metaphase progression. completion mitotic timely regulating the in WDR62 knockdown for delays Thus, requirement 3F). WDR62 (Fig. observed of was controls to compared 5100 Fg A.Qattto ftemtpaecl population cells cell WDR62-depleted of metaphase half the approximately of that spindles Quantitation indicated spindle metaphase the from 4A). abnormal centrosomes (Fig. of Strikingly, displacement in a 4A). by (Fig. resulted characterized cells depletion WDR62-depleted WDR62 and control both ora fCl cec 2 (21) 125 Science Cell of Journal a and - c tblnlclztos eeosre for observed were localizations) -tubulin 00.Hwvr h otcladsidepl oaiainof localization p150 pole spindle and and cortical Cleveland, NUMA the and pole However, (Radulescu for microtubules dynein/dynactin respectively 2010). critical astral positioning, are of and controls spindle cortex capture cortical for cell NUMA the to the and and activities of compared focusing poles spindle localization cells at motors The WDR62-depleted from in 4E). rotation in rotation (Fig. spindle of of z-axis angle degree more the greater the was a of z-axis showed z-axis measurements z-stacks orientation the the our planar the in and through in sectioning spindle 4D) (Fig. orthogonal the rotation through of seen rotation substantial clearly The WDR62- exhibited 4C). in (Fig. axis of cells polarity surface spindle to a metaphase depleted parallel the attain plane dish, addition, commonly culture division In the cells the with cultured 4B). orientation, (Fig. in planar spindle spindles metaphase bipolar while abnormal an exhibited Glued muoltigfrcci 1adcdc25C ( and phosphorylation. by B1 determined cyclin was for indicated) immunoblotting as h (2–14 thymidine release following G1/S progression at cycle synchronized Cell were (DTB). siRNA Con or WDR62 ( controls. as blotted and WDR62. siRNA) for (no immunoblotted siRNA or with siRNA) treated WDR62 (Con not a siRNA t4), non-targeting and pool, t3 siRNA t2, (t1, siRNA WDR62- targeting human individual with transfected mitotic perturbs progression. depletion WDR62 3. Fig. he needn experiments. independent means three by are identified phase Values M staining. in DAPI cells of proportion the and ( mean indicate and a values lines on Horizontal shown plot. are scatter observed, vertical were cells until daughter breakdown two envelope nuclear cells, from HeLa measured control and WDR62-depleted ( in plate. mitotis metaphase the indicate of lines position dotted the White shown. are post mitotic progression h of images 7 Representative from release. intervals thymidine 6-min at phase- optics under contrast were imaged siRNA, and WDR62 (DTB) or synchronized siRNA Con with treated ( knockdown. levels confirm WDR62 to and determined loading protein for phospho-cdc25C. blotted and B1 immunoblotting cyclin by for determined was h indicated) (0.5–10 as release h]. NZ 16 following nM, progression 350 Mitotic M- treated; at (NZ) synchronized [nocodazole were phase siRNA Con or WDR62 he needn xeiet (* experiments independent three F D6 a eltdi eaclsfr4 h 48 for cells HeLa in depleted was WDR62 ) a nlee yWR2loss WDR62 by unaltered was n ausaetettlclscutdfrom counted cells total the are values ( A eaclswr transiently were cells HeLa ) C eaclstasetdwith transfected cells HeLa ) B eaclstasetdwith transfected cells HeLa ) a and - P 6 , c D a E 0.01). ...from s.e.m. tblnwere -tubulin tblnwas -tubulin uainof Duration ) eacells, HeLa ) Journal of Cell Science irtbl yaisa otiuigfco otereduced the to with factor Furthermore, cells. contributing WDR62-depleted in a integrity indicatingcentrosome as 5F,G) (Fig. dynamics Taxol microtubule or the nocodazole in low-dose reduced of was fragmentation presence PCM of extent 5D,E). of the (Fig. Furthermore, integrity centrosome PCM reduced of suggestive disrupted was which and with co-localized that loose in centrosomes of pericentrin displaced revealed loss for a Staining cells be loss. WDR62-depleted to WDR62 appear with not cohesion did centriole there Thus, displaced 5C). in intact (Fig. be centrosomes to these found GFP- we expression, by centrin1 visualized tagged pairs, observed centriole investigated we we centrosomes Strikingly, When arrows). 5A). (Fig. apolar pole spindle multiple microtubule a in retained that and to pole spindles attachment spindle revealed the metaphase to juxtaposed abnormal remained cells of interrogation WDR62-depleted closer Our are loss poles WDR62 spindle with to compromised coupling and pole division. integrity spindle Centrosome cell for astral maintaining axis polarity or of a establish stabilizing to capture intact. spindle in integrity for role largely WDR62 cortical specific metaphase appeared localized a the highlight positioning results our and spindle Rather, for focusing microtubules in pole involved mechanisms Thus, spindle S3). Fig. material (supplementary c tblnpnt nWR2dpee el Fg 5A,B, (Fig. cells WDR62-depleted in puncta -tubulin c tblnpnt and puncta -tubulin ogeso n lnae pnl egh hs D6 is WDR62 Thus, architecture. length. reduced spindle metaphase spindle maintain with to chromosome elongated required inefficient centrosomes with and associated displaced congression also was by This integrity. metaphase typified were loss during WDR62 following defects Our spindle centrosome perturbed. show overtly studies Thus, be not to S4D). appeared cohesion Fig. and duplication material to (supplementary compared when in controls G2 numbers late centrosome at WDR62 synchronized interphase cells spindles in WDR62-depleted 2009). differences multipolar we observe However, al., not of S4C). did Fig. frequency material et supplementary 3%, the versus Silk (15% increased 2009; determination also centrosome length Mitchison, depletion spindle proper and and between pole (Dumont spindle relationship the to the attachment of recent the reinforces appreciation This control S4B). to Fig. relative WDR62-depleted material increased (supplementary significantly cells in be to length this found spindle and measured cells also we coupling, organization. spindle centrosome/bipolar the disrupted progressionand at mitotic (supplementary delayed with aligned consistent knockdown was not S4A), Fig. WDR62 chromosomes material following with plate cells metaphase of proportion increase the substantial in a demonstrating analysis, quantitative 5A– Our chromosomal (Fig. E). unaligned metaphase during of cells WDR62-siRNA-treated presence in the DNA noted we staining DAPI ihorosrain fdfcsi centrosome–spindle-pole in defects of observations our With D6 ucini ioi 5101 mitosis in function WDR62 lc ln pnl oaiyai viewed axis polarity spindle along slice (* experiments independent two from n means plot. depict scatter lines vertical Horizontal a on plotted and cells in HeLa Con-siRNA-treated measured or was WDR62- rotation spindle of ( angle orientation. planar from rotation ‘ z-axis. in rotation spindle reveals z-sections of immunostained perspective side-on a from l cl as 20 bars: scale All z-stacks. of projections show intensity images maximum Overlay cells. 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In (Fig. failed 8F). full- Fig. to the GFP–WDR62-C (GFP–WDR62-FL; compared efficiency to GFP–WDR62 reduced localization length with albeit GFP–WDR62-N spindle observed mitotic GFP– expressed non-aggregate we with ectopically cells the WDR62-N, in of However, aggregate. repeats propensity to WD40 the protein containing for region S7A,B). responsible GFP– N-terminal was Fig. while WDR62 material the despite (supplementary aggregates Thus, cytoplasmic levels distribution with expression predominantly greatly higher was cells cytoplasmic a expression to of WDR62-C led diffuse proportion mutant truncation of enhanced retained proportion WDR62-N expression Interestingly, GFP-labeled greater S7A). the that a Fig. material displayed we (supplementary cells which spindle cells, HeLa cells of the in Ad293 expressed GFP of compared analyzed aggregation WDR62 as with full-length difficulties then expressed to GFP-labeled mutants due We However, WDR62 proteins. fusion of 8D,E). in (Fig. properties expressed localization ectopically cells when JNK HeLa with interaction abrogated elsl ogtt dniytecnrbto fJNK-mediated of contribution the identify to sought lastly We ora fCl cec 2 (21) 125 Science Cell of Journal nvitro in and nvivo in nvitro in tde r ossetin consistent are studies n completely and oaiain(eatee l,20;Fn ta. 08 Varmark 2008; al., et Fong cdk5rap2, 2004; their In in al., microcephalin, centrosomal 2010). et principally (Delattre al., proteins, are localization et STIL and MCPH Pulvers CEP152, 2010; CENPJ, other by al., shared et related contrast, (Higgins is microcephaly The pole (ASPM) spindle WDR62 abnormal 2009). gene spindle of protein, Woods, localization of MCPH and another pole role (Thornton spindle the neurogenesis prominent highlights in centrosome- and integrity with proteins MCPH of associated association the reinforces functions 2010). al., et centrosome- Yu 2010; that al., the notion et underlie of the may manifestation neurogenesis al., with clinical during et defects compatible mitotic (Lesage is associated system germinal nervous This the embryonic within 2010). the self-renewal their of for compartment property neuroprogenitors, necessary of a a division as asymmetric for suggested the determinant been critical has in Indeed, orientation defects neurogenesis. cells spindle WDR62- cortical centrosome-directed that following on precursor reported orientation is specifically impacts spindle possibility neuronal depeletion and/or previously One mitosis mitotic of 2010). with timing in al., et consistent WDR62 (Nicholas of brains expression embryonic in ioi.Seiial,w eelddfcswt centrosome– with WDR62. defects lacking during cells WDR62 revealed in localized coupling spindle we spindle-pole of were loss Specifically, the cells mitosis. Thus, of knockdown result localization. and WDR62 the spindle likely in metaphase prominent defects during most centrosome/spindle its specifically with observed coincided depletion was WDR62 during the as However, phenotype structure protein. functions centrosomal WDR62 or integral exclude an number not completely did cannot centrosome depletion We al., its interphase. on et and Sir impact protein 2012; centrosome obviously a al., WDR62 that principally et demonstrated not have Hussain we is 2010; 2012), Delattre al., al., characterized et 2010; et Vulprecht well al. 2011; Hatch et 2004; with (Barrera al., CENPJ, Cep63) duplication et centriole (cdk5rap2, and in proteins Cep135 functions al. et MCPH Cep152, and Lizarraga 2010; unlike STIL, proliferation al., However, cell et in (Buchman 2010). associated reduced exit division cycle with also with cell enhanced brains neuroprogenitor latter dwarfism of mouse altered the embryonic primordial dysfunction with similarly of to pericentrin, microcephaly, and/or depletion linked or The disruption cdk5rap2 genetically 2009). proteins Woods, the PCM and involve (Thornton centrosomes also commonly may most are that proteins MCPH. these in links mutated represent that may mechanism positioning suggest plane a ASPM cleavage and and spindle WDR62 mitotic of that 2006). phenotypes al., depletion et cleavage shared (Fish altered The neuroprogenitors to similar dividing due in exit embryonic orientation cycle revealed in plane cell ASPM premature positioning of has in loss resulted spindle the mice cells Moreover, and 2010). al., cultured organization et (Higgins spindle in for The ASPM a cells. requirements Woods, human as of cultured and defects in depletion orientation (Thornton depletion and WDR62 spindle respectively of demonstrated ASPM consequence cases, for have Interestingly, of accounting We MCPH 2006). 2009). 10% in al., and mutated et frequently 50% most Zhong are 2007; WDR62 al., et u nlsso D6 nrclua itiuinadcellular and distribution intracellular WDR62 of analysis Our h oeua ai htudristedvlpeto MCPH of development the underlies that basis molecular The WDR62 uain nMP (Nicholas MCPH in mutations Journal of Cell Science rps httesidelclzto fWR2i eurdto centrosome required coupling, is WDR62 centrosome–spindle-pole of tight localization We spindle maintain proteins. the pole that spindle locations in propose maintenance characterized spindle previously mechanisms differences metaphase to mitotic different in compared the function reflect several Thus, WDR62 may underlying 2010). at defects Cleveland, spindle/centrosome functions and motors dynein/dynactin (Radulescu and 2005; multiple NUMA Scholey, Notably, and perform 2009). spindle Morales-Mulia inactivated al., 2010; of et al., centrosome Silk loss have reduced et that to (Barr lead be shown that necessarily integrity not have that does may studies dynein attachment pole or events suggests previous NUMA fragmentation which cdk5rap2, with However, and dynamics, related. As uncoupling microtubule centrosome knockdown. formation centrosome spindle required in bipolar WDR62 to and defects subsequent observed to uncoupling, were integrity response pole centrosome/spindle in astral centrosome or reduced maturation suggest integrity centrosome findings in our defects formation, find microtubule not size did a we in such as smaller to and were lead centrosomes also disrupted may pole MT However, spindle to phenotype. uncoupled recruitment positively the PCM on that puncta ends possible minus is multiple It pericentrin. with for stained diffuse appeared spindle–centrosome centrosomes for of spindle critical mislocalization the and factors attachment. by identified bipolarity explained readily forces previously spindle not microtubule but establishing to orientation with linked of disruption is associated the that coupling suggests pole by therefore centrosome–spindle unaltered spindle study were Our the dynein loss. and addition, WDR62 NUMA not In of underlie localization did poles. cortical spindle may and depletion unfocused WDR62 forces in result However, microtubule that uncoupling. that noted centrosome we spindle suggesting Interestingly, bipolar to 2009). formation subsequent al., occurred and displacement et Morales-Mulia centrosome Silk 2010; 2005; al., et Scholey, or (Barr NUMA subunits cdk5rap2, centrosomal dynein/dynactin of disruption the with observed opnn fteJKsgaigptwy(asra tal., a during et maintained (Wasserman is as this pathway that demonstrate characterized signaling findings our JNK and initially 2010) the of was component WDR62 phosphorylation. more regulation. functional appears in involved phosphorylation be to WDR62 likely spindle Rather, mitotic WDR62, for WDR62 sufficient of localization. is region regulating that sites, N-terminal phosphorylation in the C-terminal entry, this, indicates lacking involved of mitotic support likely This In movement. to not However, phosphorylation. is prior phosphorylated. phosphorylation WDR62 centrosomes, its mitotic mitotically to regulatory preceded regulate first movement is of that provided WDR62 range now WDR62 mechanisms has the the study activity. 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(DTB) 10 synchronized were CaCl and cells inhibition HeLa Eg5 assays, regrowth Microtubule and (10 ng), substrate (10 protein recombinant Purified vitro In arr,J . a,L . amr .E,Sean . uh,J .adMegraw, and L. J. Fuchs, J., Seemann, E., R. Hammer, R., L. Kao, F. A., J. Gergely, Barrera, and V. J. Kilmartin, R., A. Barr, Sillje A., F. Barr, F. Gergely, and R. A. Barr, References respectively.]. at M.A.B., http://jcs.biologists.org/lookup/suppl/doi:10.1242/jcs.107326/-/DC1 online available and material Supplementary months. D.C.H.N 6 after release to for National PMC 566804 in and Deposited numbers and [grant J.I.H.]; Grants Project to 628335 Council Research APP1011505 Medical an and number Health D.C.H.N.]; Development and Career [grant [to Council Dentistry Fellowship Research Fellowship Medicine, Medical Roper and of CR Health Faculty National (MDHS) a Science by supported Health was work This manuscript. Funding this of reading Biology/South critical Tilley Cancer for Leann Melbourne) Prof. for and of (Centre antibody (University NEDD1 Kumar the for Sharad Pathology) Australia Prof. thank We Acknowledgements counted exclusion. were Blue numbers Trypan Cell and depletion. hemocytometer WDR62 confirm a levels the to using protein parallel WDR62 using in specifications. blotted manufacturer hydrate}, were to acid according (Roche) sulfonic Kit XTT benzene (4-methoxy-6-nitro) tetrazolium]-bis tie ihGloeBu ti egn n nlzdb uoaigah and autoradiography by analyzed and SDS-PAGE, reagent by Stain resolved counting. then Blue Cerenkov were Gelcode Samples with buffer. stained sample Laemmli of addition eelkntcoemcouue,snhoie el eeetatdwt Calcium with CaCl extracted mM were (1 Buffer cells Extraction synchronized microtubules, kinetochore reveal 37 iula agns ope farlmd-edn hstglgn (Mn ligand Phos-tag acrylamide-pendant of complex manganese dinuclear ecinbfe 2 MHPSp .,2 MMgCl mM 20 7.6, pH HEPES mM (20 buffer reaction 0mM 20 T)oe 2 i iecus t30 at course time min 120 a over DTT) a;50 tag; / rtnX10 o i eoefxn n muoloecneanalysis. immunofluorescence and fixing before min 3 for X-100) Triton v/v ˚ 114. J. Biochem. A. Aronheim, malformations. brain severe in mutations 207-210. WDR62 recessive identifies Tu D., d5a2itrcswt eieti omiti h erlpoeio oli the in pool progenitor neural the maintain to neocortex. pericentrin developing with interacts Cdk5rap2 Cell .L. T. response. damage DNA and attachment 23-39. pole spindle to centrosome fcl division. cell of Sci. Cell J. o ewe n 0mnbfr iain o iei g niiin HeLa inhibition, Eg5 kinesin For fixation. before min 60 and 2 between for C vr . Oztu K., ¨var, 18 ¨ysu m 21) D5A2rgltscnroeeggmn n oeini mice. in cohesion and engagement centriole regulates CDK5RAP2 (2010). b iaeassay kinase M). 913-926. , gyeohsht n upeetdwt 25 with supplemented and -glycerophosphate z . Cag B., ¨z, ˚ ,3 i) irtbl erwhwsiiitdb nuaigclsat cells incubating by initiated was regrowth Microtubule min). 30 C, m 120 439 32 )wr rae with treated were g) ,H .adNg,E A. E. Nigg, and H. 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