Phosphorylation network dynamics in the control of transitions Daniel Fisher, Liliana Krasinska, Damien Coudreuse, Béla Novák

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Daniel Fisher, Liliana Krasinska, Damien Coudreuse, Béla Novák. Phosphorylation network dynamics in the control of cell cycle transitions. Journal of Cell Science, Company of Biologists, 2012, 125 (Pt 20), pp.4703–4711. ￿10.1242/jcs.106351￿. ￿hal-01068356￿

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Journal of Cell Science o:10.1242/jcs.106351 doi: 4703–4711 125, Science Cell of Journal 1 Fisher Daniel of transitions control cycle the cell in dynamics network Phosphorylation Commentary 04.I diin d1i niie hnbudt d inhibitory p21 (in Cdk the to bound including when (CKIs), inhibited (Murray, proteins is cycle Cdk1 cell the addition, anaphase during In stages 2004). ligase specific at activity ubiquitin regulated Cdk1 the of the for loss crucial by is (APC/C), initiated Cyclin complex/cyclosome the promoting 2001). of is al., affinity which et the (Wohlschlegel modulates 2004), degradation, (Russo substrates also (CAK)-mediated its subunit loop Roberts, towards cyclin complex kinase-activation The kinase and 1996). conserved al., Cdk-activating (Sherr et its cycle of through sequentially phosphorylation are is cell directly that Cdk1 the cyclins 1). and several (Fig. during of other one each expressed binding with by interact activated also that components Santamarı Hochegger 2008a; 2009); al., 2007; et al., Krasinska al., et and 2008; et replication al., (Malumbres DNA et (Hochegger orderly cells segregation ensure to eukaryotic chromosome sufficient been the is all far, for it essential in so moreover, is that have, cycle one only that other the DNA cell many Cdks is in Cdk1 and twenty humans, roles in the have identified also Of triggering to processes. for known cellular now essential are identified are but originally replication that were Cdks kinases activity. regulatory as their a for of require subunit family that cyclin a kinases constitute serine/threonine (Cdks) heterodimeric kinases protein Cyclin-dependent Introduction transitions. dynamic ensuring in of the role model essential quantitative underlie an general a have words: that to activities Key lead phosphatase processes considerations of and These levels gradual. biochemical kinase directionality threshold than generates opposing the rather activity the which abrupt Cdk how in are in to consider on transitions increase control, order cycle light an cycle must cell in how shed cell our why determined, cycle have are and we redefined cell that phase cycle, each cell the findings question, has through the through recent progression in model progression this discuss for required ordered This we are answer generate Commentary, that mitosis. kinase activity this To Cdk1 into one In of substrates. entry mitosis? to activity Cdk1 applicable promotes be from the of to phosphorylation it in appear replication change threshold, now principles a DNA specific main does the its a triggers separate how and Cdk1 progression, But exceeds low, cycle cells. is cell protein level activity eukaryotic organize single its that a activity all When inputs of (Cdk1). functional activity its essential 1 the kinase when the in dependent changes of phase; cyclin quantitative understanding and by S driven – be B of can cyclin yeast namely onset fission – in cyclin cycle a cell comprising the complex that kinase proposed was it ago, years Fifteen Summary ( ` correspondence for *Author 3 2 he d2 sfrsi etbae Cc5,Cc5 and Cdc25B (Cdc25A, vertebrates the in Of this phosphatases. isoforms 1992); Cdc25 al., Cdc25 the et by three (Parker by counteracted inactive Tyr15 is becomes on modification Cdk1 phosphorylated kinase, when phosphorylation: tyrosine by hs uhr otiue qal oti work this to equally contributed authors These xodCnr o nertv ytm ilg,Dprmn fBohmsr,Uiest fOfr,SuhPrsRa,Ofr X Q,UK 3QU, OX1 Oxford Road, Parks South Oxford, of University France Biochemistry, Rennes, of 35043 Department 6290, Biology, UMR Systems CNRS Integrative Rennes, for of Centre Development Oxford and Genetics of Institute Ge de Institut 02 ulse yTeCmayo ilgssLtd Biologists of Company The by Published 2012. euaino d1atvt novssvrlseparate several involves activity Cdk1 of Regulation Cip1 aiy(nvrerts.Ck ucini locontrolled also is function Cdk1 vertebrates). (in family PA elcce ylndpnetkns,Popaae uniaiemodel Quantitative Phosphatase, kinase, Cyclin-dependent cycle, Cell PP2A, ´ne tqeMole ´tique 1, ,LlaaKrasinska Liliana *, [email protected] clied otele,IM,CR M 55 Universite 5535, UMR CNRS IGMM, Montpellier, de ´culaire .cerevisiae S. ) 1, ` ainCoudreuse Damien , ´ n ebr of members and ) ta. 2007). al., et a hs xrm ipiiysilapasvldtdy o xml,in example, For today. a 2). valid appears (Fig. still exceeds simplicity triggered extreme whose is activity onset mitotic Cdk1 threshold, once – intermediate high Finally, G1 an – attains DNA specific 2004). in activity bulk (Diffley, Cdk1 by origins once followed level is phase, are replication This S low. in DNA events be synthesis and to of (Fisher activity cycle Cdk1 activity Licensing requires Cdk1 cell 1996). in changes different Nurse, by whereby triggered cell of sequentially model’ This regulation, ‘quantitative 1996). the Nurse, of cycle and proposal (Fisher the mitosis to and led observation phase the S promote both can of complex model cyclin-B–Cdk1 onset quantitative single a the yeast, – fission Cdk1 In by control cycle Cell eteo ewr frgltr nye,woeoutput whose enzymes, regulatory the at of activity responses. network system-wide Cdk1 coherent a generates counteract but of different that Cdk1 place centre the phosphatases only general not does through those simple model activity This also progression cycle. a Cdk1 cell ordered the in propose of mediate phases variations and how can from explain cycle, alone could studies quantitative cell that a on the with hypothesis draw compatible of are poorly we that regulation cell remain systems Commentary, the model transitions this multiple control unidirectional activity In Cdk1 promoting understood. in by changes in cycle resulting way the the and networks which phosphorylation these of characteristics abruptly is and Nilsson Cdc25) 2005; al., as 2000). et (Ferguson Hoffmann, to transition G2–M the referred at activated (hereafter Cdc25C Cdc25C), 2, ` oercn tde aeln diinlspott hsmodel, this to support additional lent have studies recent More ncnrs oteewl-ecie ehnss h dynamic the mechanisms, well-described these to contrast In n Be and otele n I 49 otele,France Montpellier, 34293 II, and I Montpellier ´ l Nova ´la ´k 3, ` 4703 Journal of Cell Science eo h hehl oalwrpiainlicensing). replication drop allow must not to activity do threshold (kinase mitosis maximal the into or below activity) entry kinase or stage. insufficient onset cycle with (S-phase occur cell minimal the either determine are activity) quantitative Thresholds descending the or ascending – (i.e. activity history Cdk changing by control model. cycle Cell 2. Fig. be might cycle. cell cyclin entire the mitotic the through single yeast that budding a possible drive to therefore, of sufficient seems, pattern It expression cyclins 2005). correct mitotic Aparicio, and – S-phase conditions and the certain (Hu under of functions – essential can Clb2, all Clb2 Clb1, perform mitotic cyclins the mitotic Clb4, the and and S- Clb3 the Clb6, Cln3, and and Clb5 Cln2 in cyclins Cln1, essential phase cyclins G1 are the cyclins namely yeast yeast, of budding families fission three and Although the respectively). budding is between But ( similarities 2010). There certainly, Nurse, organisms? all are, and to applicable (Coudreuse universally events model quantitative inappropriate order cycle the change Moreover, cell even can 2011). complex of this Novak, of activity the and of wild-type alteration the (Tyson recapitulate cycle can modelling system cell mathematical this Recently, how sequentially demonstrated mitosis. cell has and a phase the of activity S drives the through combination of modulation activity cyclin-B–Cdk1 simple by Cdk1 the single words, governed that other Nurse, and In artificially so (Coudreuse 2010). thresholds be high inhibitor and low can of between oscillates concentration cycle the cell changing entire passage the the inhibitor, small-molecule non- a through to and sensitive uniquely active be engineered constitutively to been has a that protein express fusion gi cyclin-B–Cdk1 that degradable are homologues cells functional yeast Probable regulators. text. fission additional the other to and refer lines) please straight details, by more (indicated For activity colour. of cycle. same cell windows the respective during their regulation phosphatase humans, and Cdk 1. 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Cdk1 represents axis vertical The ora fCl cec 2 (20) 125 Science Cell of Journal Min Min H. sapiens S. cerevisiae S phase PP2A–B55 Cip/Kip and Sic1 Mitosis Cdc28–Cln1,2 cioacaoye pombe Schizosaccharomyces Cdk activity Cdk2–cycE omtetpit/Sat 2M Metaphase/Anaphase G2–M Commitment point/Start G1 Cdc28–Clb5,6 Licensing Cdk2–cycA Max Cdc28–Clb3,4 oprsno d opee n hshtssta oto h elccei udn es and yeast budding in cycle cell the control that phosphatases and complexes Cdk of Comparison S , Cdc28–Clb1,2 Cdk1–cycB loudriscl yl euaini eaon.I ie for mice, In metazoans. in regulation cycle cell underlies also osblt sta hscmlxt rvdsrbsns by robustness provides complexity this are why that order, correct is the One in date? to possibility mitosis studied eukaryotes and all in phase involved complexes multiple S promote the to of – result a as indispensable a them. single become between in might a interactions but them Thus, of sufficient, E. several Cdk2 minimally and because be A cycle normal probably might cyclins cell 2012), complex S-phase in cyclin–Cdk al., nuclear and role et the 2011) al., rate-limiting titrates (Echalier et a extracts (Merrick cells has egg – human Cdk2 that in shown progression – have present approaches genetics when explanation). chemical for at example, 2008b regulators al., For all et of Krasinska (see presence levels the the physiological as in not inhibited conclusions are reflect transitions. might enzymes same specific experiments cycle the not cyclins knockdown to cell and do lead or important Cdks depletion obviously these multiple Furthermore, they promote which in mitosis, successively situation, and controlling physiological in phase redundancy cyclin–Cdk S of principles the revealed cyclin- single a with B operate cyclin complex. to or Cdk cells is A vertebrate protein cyclin allow of it either even timing of might considerations, the levels and in these localisation manipulation expression, Given that speculate which transitions. to both 2003), tempting drive cycle to al., capacity cell intrinsic et the main (Moore has B replication cyclin that DNA demonstrates promote in to nucleus the able to relocalised when is Strikingly, S-phase B 2011). both cyclin al., to (Strausfeld et bind (Pagliuca mitosis can substrates A and Cdk cyclin mitotic phase cells, and human S in both and 1996), of al., onset et the promote can A ( genes cells E most cyclin in both cycle (Santamarı cell development the early control during to sufficient is Cdk1 example, hs Klscysae l,20) In S 2009). promoting al., for et A (Kalaszczynska cyclin phase with redundant are and progression cycle ubro eot aesgetdta h uniaiemodel quantitative the that suggested have reports of number A oevr fasnl ylnCkcmlxsfie nprinciple in – suffices complex cyclin–Cdk single a if Moreover, have situations experimental various these although However, Inactive Cdk1–cycB nvivo in G2 P iuto,we ait fcmlxsaepresent, are complexes of variety a when situation, Cdc25 Wee1

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White, M., A. Sheinerman, Ferguson, P., Jay, F., Hoh, E., Hodimont, A., Camasses, E., Cot, A., Echalier, u .adAaii,O M. O. Aparicio, and F. Hu, T. Hunt, and S. Takeda, H., Hochegger, oawl,M,d rzeSla .F,Hl,F . ilas .T,Carbonaro-Hall, T., R. Williams, L., F. Hall, F., E. Silva, e Cruz da M., Dohadwala, F. J. Diffley, ohge,H,Djuhn,D,Snd,E,Sbr,A,Rjnr,E,Kr,J,Hunt, J., Kirk, E., Rajendra, A., Saberi, E., Sonoda, D., Dejsuphong, H., Hochegger, ioht,N,Ymn,H,Nw,H,Ysia .adYngd,M. Yanagida, and T. Yoshida, H., Niwa, H., Jr Yamano, E., N., J. Kinoshita, Ferrell, and Y. S. Kim, Silver, I., Kondratiuk, Y., Choi, S., Mizuno, T., Iino, Y., Geng, I., Kalaszczynska, ageo osiln ..i upre yE P (MitoSys). FP7 EU by supported is B.N. Roussillon. Languedoc odes,D n us,P. Nurse, and D. Coudreuse, avy .L,Ecs,G,Dpor,N,Gg,S . uaadn,J n Kellogg, and J. Gunawardena, P., S. Gygi, N., Dephoure, G., Enciso, L., S. Harvey, J. Gunawardena, lre .R,Hfmn,I,Data .adKret,E. Karsenti, and G. Draetta, I., Hoffmann, R., P. Clarke, Labbe E., Brioudes, S., Vigneron, A., Burgess, F. Uhlmann, and C. Re Bouchoux, the References by supported also is lab long-term HFSPO The an by (LT-00623). supported an fellowship is is D.C. L.K. and fellow. EL2010.LNCC/DF. ANR-09-BLAN-0252-01 postdoctoral Cancer, ARC le grant Contre ANR Nationale by Ligue the funded was work This manuscript. the Funding on comments critical for Dulic Vjeko thank We Acknowledgements 4710 rr,C n odee,A. Goldbeter, and C. ´rard, rr,C,Gne .adGlbtr A. Goldbeter, and D. Gonze, C., ´rard, oaetmdfcto nbooia systems. biological in modification covalent 2A. phosphatase protein inhibiting by 1673-1677. mitosis controls T. Lorca, Arpp19, and kinase, A. Castro, A., Van-Dorsselaer, E., h outeso siltosi h ewr fcci-eedn iae rvn the driving kinases cyclin-dependent of network cycle. the cell in mammalian oscillations of robustness the cycle. cell 21648. mammalian the driving network mitosis. to entry cyclins. G1 of absence the in mitosis 850-860. and S-phase both promotes kinase 20) lblaayi fCk usrt hshrlto ie rvdsisgt into insights provides sites phosphorylation substrate evolution. Cdk1 of analysis Global (2009). omlcl yl n hcpitrsossi ieadclslcigCc5 and oscillate? Cdc25B circuits lacking certain cells and mice in responses phosphatases. checkpoint protein Cdc25C and cycle cell sensitivity. Normal inhibitor and redundancy functional cdk2 Biol. into insight D. Fisher, provides and L. Krasinska, B., F. yl rniin:de n i all? fit one does transitions: cycle .A,Nin .C,Genad .adBrd,N. kinases. Berndt, cyclin-dependent and by USA P. 1 Sci. phosphatase Greengard, protein C., of A. inactivation Nairn, A., D. 14 eaierglto fmtssb h iso es rti hshts ppa2. phosphatase protein yeast fission the by Dev. mitosis of regulation Negative Wee1. of inactivation the in ultrasensitivity cells. stem embryonic Saccharomyces and P. hematopoietic 365. in Sicinski, in essential and but K. proteins fibroblasts Akashi, in J., D. replication Wolgemuth, P., toward D. cyclins mitotic of cerevisiae. activity the cells. vertebrate in genetics chemical via S. Takeda, and T. oto network. control nbdigyeast. budding in R. D. 6844. fcc5Cb ye2 rti hshts:seii euaindrn h elcycle cell the during extracts. regulation egg specific phosphatase: Xenopus protein in type-2A a by cdc25-C of due defects balance. mitotic multiple B-Cdc2/PP2A and cyclin 12564-12569. arrest the G2 of in deregulation results to Greatwall human of Loss (2010). exit. mitotic during dephosphorylation a eapo switch. poor a be can R778-R786. , 7 19 21) hshts hehl estelvlo d1atvt nerymitosis early in activity Cdk1 of level the sets threshold phosphatase A (2011). 1059-1071. , 1028-1040. , 91 ora fCl cec 2 (20) 125 Science Cell of Journal Science 20) euaino al vnsi hoooereplication. chromosome in events early of Regulation (2004). rc al cd c.USA Sci. Acad. Natl. Proc. 6408-6412. , 20) utst rti hshrlto ae odtrsodbut threshold good a makes phosphorylation protein Multisite (2005). e.Cell Dev. Nature o.Bo.Cell Biol. Mol. 325 20) nesnilrl o d1i hs oto srevealed is control phase S in Cdk1 for role essential An (2007). rc al cd c.USA Sci. Acad. 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Xenopus in control phase ml oeue orva eurmnsi ua elproliferation. cell P. human in R. requirements Fisher, reveal and to molecules M. small K. Shokat, A., Goga, 4843-4852. td d1adCk ucin ntecl cycle. cell the in functions Cdk2 and Cdk1 study .L,Suebr,P .adGrsy .J. cells. G. Gorbsky, vertebrate and in T. P. Stukenberg, L., D. 351. family. fDAplmrs lh nteiiito fDAreplication. DNA of initiation binding the chromatin for in required alpha is polymerase RecQ4 DNA Xenopus of of region noncatalytic N-terminal oocytes. frog of maturation meiotic Cdc25. of activation S. feedback Kornbluth, positive and A. T. elcto rgnfrn nXenopus. in firing origin replication D. Fisher, esatrsodfrteosto N replication. DNA of onset the for threshold M. a Tyers, sets and T. Pawson, F., d2 dephosphorylation. J. Cdc25 D. Wolgemuth, S. Kornbluth, and H. L. portrait. family Tsai, a kinases: O., D. feedback. Morgan, systems-level of consequence the is exit mitotic i1dsrcina h ne fSphase. 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