Cdt1

 

Loads Mcm2-7 complex on DNA at  at ORC in pre-RC/licensing step. Inhibited by geminin.

 

Synonyms

Tah11; Sid2; RLF-B; Dup

Molecular weight

68 kDa (Sc)

Biochemical properties

• Binds DNA (1)

• role in preIC formation with cdc7 {Ballabeni et al., 2009, J Biol Chem, 284, 3028-36}

Motifs

•PIP box (2) (3)

Protein interactions

Sc

•Mcm27 (4) (5)

•topoI/sicI (6)

 

Sp

•Cdc18 (7)

•Mcm27 (8)

 

Xl

•Geminin (9) stoichiometry of cdt1:geminin in complex determines activity (10)

•Cdt2 (11)

•Ddb1 (2)

•PCNA (2)

• MCM9 {Lutzmann and Mechali, 2008, Mol Cell, 31, 190-200}

 

Dm

•Cdk (12)

•Mcm10 (13)

 

Mammals

•Geminin (14) (15) (1) 70% cell cdt1 bound to geminin (16), affected by polyunsaturated fats and co enzyme Q10 {Mizushina et al., 2008, Int J Mol Med, 21, 281-90} {Mizushina et al., 2008, Biochim Biophys Acta, 1780, 203-13}, and protein X hep B {Rakotomalala et al., 2008, J Biol Chem, 283, 28729-40}

•Mcm2-7 (1) mouse {Teer and Dutta, 2008, J Biol Chem, 283, 6817-25} human.

•ORC2 (1)

•Cyclin A-CDK (17)

•Cdt2/L2DTL (18)

•PCNA (3)

•Cdc6 (19)

•cdc7:dbf4

{Ballabeni et al., 2009, J Biol Chem, 284, 3028-36}

• screen – mcm4/6, apc,snf2,wstf,pcna,topoI/II,importin alpha and beta , nucleophospmin,grwd1, histones {Sugimoto et al., 2008, Mol Biol Cell, 19, 1007-21}

• HBO1 acetylase {Miotto and Struhl, 2008, Genes Dev, 22, 2633-8}

 

Plants

• GEM (plant geminin) {Castellano Mdel et al., 2004, Plant Cell, 16, 2380-93}

Zebra fish

• cdt2 (20)

 

Modifications

•Phosphorylation:

Dm at least 10 sites (12)

Mammals (17) (21)

Structure

•With geminin fragment (22)

•On sizing columns cdt1 seems to be pentamer/ hexamer –ring around the DNA (23)

 

Cellular location and expression

Sc

• protein levels very little cell cycle variation (6) (4)

•nuclear M-G1 then cytoplasm (6) (4)

 

Pombe

•some control of expression transcriptional (8) via cdc10 (24)

•Do see chromatin loading – independent cdc6 (7) peaks in G1 (just before mcms) (8)

•Proteolyzed during S phase using Cul4-Ddb1-Cdt2 ubiquitin ligase (25)

 

Xenopus

•Inhibited by geminin (9) (14)

• in vitro associates with chromatin at the same time as mcm4 before cdc45 loads (26) but must bind after cdc6 to get functionality (27)  Leaves as replication starts. See also (16)

•Geminin and cdt1 bind to chromatin at the same time (28)

• Unstable protein degraded in mitosis via ub and proteosome. (29) dependent on pcna (26)  (3) and Cul4-Ddb1-Cdt2 ubiquitin ligase (11). Ub forms only on the chromatin (26) suggest that this is the form that binds pcna.

• cdt2 needed for breakdown in response to MMS ( needs PCNA) (11)

 

mammals

•Inhibited by geminin

• unstable. broken down by proteosome (15) (30) part via skp2 after phosphorylation (30) other dependent pcna (21) and Cul4-Ddb1-Cdt2 ubiquitin ligase (31) (32) (33).

•Degraded on DNA damage via proteosome (34) depends on Cul4-Ddb1-Cdt2 ubiquitin ligase but not atr/atm (35)

• unstable in the absence geminin in mouse embryos (36)

• nuclear g1 , absent s on (15) not seen in cytoplasm(37)

• Low in quiescent cells (38)

• spatio-temporal dynamics {Sakaue-Sawano et al., 2008, Cell, 132, 487-98}

 

Plants

• expression patterns {Castellano Mdel et al., 2004, Plant Cell, 16, 2380-93}

 

c.elegans

•s phase degradation via ddb1 and cul4 (39), (40)

 

Other comments

•Needs ORC for chromatin binding (7) (41)

 

Sc

•Cdt1 needed in vitro for mcm to bind to bubble or y substrates. (5)

•Quite distant from other species only 12% by sequence

Sp

• Overexpression alone does not case re replication (8) (24) but does if with cdc18. (8)

 

Dm

•Role for e2f and dp1 in expression (42)

•In chorion amplification see dup at replication fork (12)

 

Xl

•Addition extra gives re replication in G2 but not M G1 S (43). At very high levels re replication decreases ( partial activation atr) (44)

 

Mammals

•Overexpression can cause re-replication in p53-deficient background (associated with an increase in DNA damage) (45)

• Geminin nuclear transport depends on cdt1 (46)

•Tumorigenic in mice if overexpressed (47)

•Long term overexpression cdt1 leads to chromosomal instability (not short term) (48)

• proteolysis regulated by cyclin D1 (49)

• levels affected by p16ink4a (50)

• Geminin depletion decreases cdt1. Suggest caused by dsbs generated by re replication rather than blocking of ub ligase by direct interaction geminin and cdt1 {Hall et al., 2008, J Biol Chem, 283, 25356-63}

• Overproduced cdt1 causes re replication in transformed cells ( much less if non transformed) {Dorn et al., 2009, Nucleic Acids Res, 37, 60-9}

Revised by

 

Last edited

9 July 09

 

 

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2.    Arias,E.E. and Walter,J.C. (2006) PCNA functions as a molecular platform to trigger Cdt1 destruction and prevent re-replication. Nat Cell Biol 8, 84-90.

3.    Senga,T., Sivaprasad,U., Zhu,W., Park,J.H., Arias,E.E., Walter,J.C. and Dutta,A. (2006) PCNA is a cofactor for Cdt1 degradation by CUL4/DDB1-mediated N-terminal ubiquitination. J Biol Chem 281, 6246-6252.

4.    Tanaka,S. and Diffley,J.F. (2002) Interdependent nuclear accumulation of budding yeast Cdt1 and Mcm2-7 during G1 phase. Nat Cell Biol 4, 198-207.

5.    Kawasaki,Y., Kim,H.D., Kojima,A., Seki,T. and Sugino,A. (2006) Reconstitution of Saccharomyces cerevisiae prereplicative complex assembly in vitro. Genes Cells 11, 745-756.

6.    Devault,A., Vallen,E.A., Yuan,T., Green,S., Bensimon,A. and Schwob,E. (2002) Identification of Tah11/Sid2 as the ortholog of the replication licensing factor Cdt1 in Saccharomyces cerevisiae. Curr Biol12, 689-694.

7.    Nishitani,H., Lygerou,Z., Nishimoto,T. and Nurse,P. (2000) The Cdt1 protein is required to license DNA for replication in fission yeast. Nature 404, 625-628.

8.    Gopalakrishnan,V., Simancek,P., Houchens,C., Snaith,H.A., Frattini,M.G., Sazer,S. and Kelly,T.J. (2001) Redundant control of rereplication in fission yeast. Proc Natl Acad Sci U S A 98, 13114-13119.

9.    Tada,S., Li,A., Maiorano,D., Mechali,M. and Blow,J.J. (2001) Repression of origin assembly in metaphase depends on inhibition of RLF-B/Cdt1 by geminin. Nat Cell Biol 3, 107-113.

10.  Lutzmann,M., Maiorano,D. and Mechali,M. (2006) A Cdt1-geminin complex licenses chromatin for DNA replication and prevents rereplication during S phase in Xenopus. EMBO J 25, 5764-5774.

11.  Jin,J., Arias,E.E., Chen,J., Harper,J.W. and Walter,J.C. (2006) A family of diverse Cul4-Ddb1-interacting proteins includes Cdt2, which is required for S phase destruction of the replication factor Cdt1. Mol Cell 23, 709-721.

12.  Thomer,M., May,N.R., Aggarwal,B.D., Kwok,G. and Calvi,B.R. (2004) Drosophila double-parked is sufficient to induce re-replication during development and is regulated by cyclin E/CDK2. Development 131, 4807-4818.

13.  Christensen,T.W. and Tye,B.K. (2003) Drosophila MCM10 interacts with members of the prereplication complex and is required for proper chromosome condensation. Mol Biol Cell 14, 2206-2215.

14.  Wohlschlegel,J.A., Dwyer,B.T., Dhar,S.K., Cvetic,C., Walter,J.C. and Dutta,A. (2000) Inhibition of eukaryotic DNA replication by geminin binding to Cdt1. Science 290, 2309-2312.

15.  Nishitani,H., Taraviras,S., Lygerou,Z. and Nishimoto,T. (2001) The human licensing factor for DNA replication Cdt1 accumulates in G1 and is destabilized after initiation of S-phase. J Biol Chem 276, 44905-44911.

16.  Xouri,G., Squire,A., Dimaki,M., Geverts,B., Verveer,P.J., Taraviras,S., Nishitani,H., Houtsmuller,A.B., Bastiaens,P.I. and Lygerou,Z. (2007) Cdt1 associates dynamically with chromatin throughout G1 and recruits Geminin onto chromatin. EMBO J 26, 1303-1314.

17.  Sugimoto,N., Tatsumi,Y., Tsurumi,T., Matsukage,A., Kiyono,T., Nishitani,H. and Fujita,M. (2004) Cdt1 phosphorylation by cyclin A-dependent kinases negatively regulates its function without affecting geminin binding. J Biol Chem 279, 19691-19697.

18.  Higa,L.A., Banks,D., Wu,M., Kobayashi,R., Sun,H. and Zhang,H. (2006) L2DTL/CDT2 interacts with the CUL4/DDB1 complex and PCNA and regulates CDT1 proteolysis in response to DNA damage. Cell Cycle 5, 1675-1680.

19.  Cook,J.G., Chasse,D.A. and Nevins,J.R. (2004) The regulated association of Cdt1 with minichromosome maintenance proteins and Cdc6 in mammalian cells. J Biol Chem 279, 9625-9633.

20.  Sansam,C.L., Shepard,J.L., Lai,K., Ianari,A., Danielian,P.S., Amsterdam,A., Hopkins,N. and Lees,J.A. (2006) DTL/CDT2 is essential for both CDT1 regulation and the early G2/M checkpoint. Genes Dev 20, 3117-3129.

21.  Takeda,D.Y., Parvin,J.D. and Dutta,A. (2005) Degradation of Cdt1 during S phase is Skp2-independent and is required for efficient progression of mammalian cells through S phase. J Biol Chem 280, 23416-23423.

22.  Lee,C., Hong,B., Choi,J.M., Kim,Y., Watanabe,S., Ishimi,Y., Enomoto,T., Tada,S., Kim,Y. and Cho,Y. (2004) Structural basis for inhibition of the replication licensing factor Cdt1 by geminin. Nature 430, 913-917.

23.  Madine,M. and Laskey,R. (2001) Geminin bans replication licence. Nat Cell Biol 3, E49-50.

24.  Hofmann,J.F. and Beach,D. (1994) cdt1 is an essential target of the Cdc10/Sct1 transcription factor: requirement for DNA replication and inhibition of mitosis. EMBO J 13, 425-434.

25.  Ralph,E., Boye,E. and Kearsey,S.E. (2006) DNA damage induces Cdt1 proteolysis in fission yeast through a pathway dependent on Cdt2 and Ddb1. EMBO Rep 7, 1134-1139.

26.  Arias,E.E. and Walter,J.C. (2005) Replication-dependent destruction of Cdt1 limits DNA replication to a single round per cell cycle in Xenopus egg extracts. Genes Dev 19, 114-126.

27.  Tsuyama,T., Tada,S., Watanabe,S., Seki,M. and Enomoto,T. (2005) Licensing for DNA replication requires a strict sequential assembly of Cdc6 and Cdt1 onto chromatin in Xenopus egg extracts. Nucleic Acids Res 33, 765-775.

28.  Maiorano,D., Rul,W. and Mechali,M. (2004) Cell cycle regulation of the licensing activity of Cdt1 in Xenopus laevis. Exp Cell Res 295, 138-149.

29.  Li,A. and Blow,J.J. (2005) Cdt1 downregulation by proteolysis and geminin inhibition prevents DNA re-replication in Xenopus. EMBO J 24, 395-404.

30.  Li,X., Zhao,Q., Liao,R., Sun,P. and Wu,X. (2003) The SCF(Skp2) ubiquitin ligase complex interacts with the human replication licensing factor Cdt1 and regulates Cdt1 degradation. J Biol Chem 278, 30854-30858.

31.  Hu,J. and Xiong,Y. (2006) An evolutionarily conserved function of proliferating cell nuclear antigen for Cdt1 degradation by the Cul4-Ddb1 ubiquitin ligase in response to DNA damage. J Biol Chem281, 3753-3756.

32.  Lovejoy,C.A., Lock,K., Yenamandra,A. and Cortez,D. (2006) DDB1 maintains genome integrity through regulation of Cdt1. Mol Cell Biol 26, 7977-7990.

33.  Nishitani,H., Sugimoto,N., Roukos,V., Nakanishi,Y., Saijo,M., Obuse,C., Tsurimoto,T., Nakayama,K.I., Nakayama,K., Fujita,M., Lygerou,Z. and Nishimoto,T. (2006) Two E3 ubiquitin ligases, SCF-Skp2 and DDB1-Cul4, target human Cdt1 for proteolysis. EMBO J 25, 1126-1136.

34.  Kondo,T., Kobayashi,M., Tanaka,J., Yokoyama,A., Suzuki,S., Kato,N., Onozawa,M., Chiba,K., Hashino,S., Imamura,M., Minami,Y., Minamino,N. and Asaka,M. (2004) Rapid degradation of Cdt1 upon UV-induced DNA damage is mediated by SCFSkp2 complex. J Biol Chem 279, 27315-27319.

35.  Blow,J.J. and Dutta,A. (2005) Preventing re-replication of chromosomal DNA. Nat Rev Mol Cell Biol 6, 476-486.

36.  Hara,K., Nakayama,K.I. and Nakayama,K. (2006) Geminin is essential for the development of preimplantation mouse embryos. Genes Cells 11, 1281-1293.

37.  Kulartz,M. and Knippers,R. (2004) The replicative regulator protein geminin on chromatin in the HeLa cell cycle. J Biol Chem279, 41686-41694.

38.  Xouri,G., Lygerou,Z., Nishitani,H., Pachnis,V., Nurse,P. and Taraviras,S. (2004) Cdt1 and geminin are down-regulated upon cell cycle exit and are over-expressed in cancer-derived cell lines. Eur J Biochem 271, 3368-3378.

39.  Kim,J., Feng,H. and Kipreos,E.T. (2007) C. elegans CUL-4 prevents rereplication by promoting the nuclear export of CDC-6 via a CKI-1-dependent pathway. Curr Biol 17, 966-972.

40.  Korzelius,J. and van den Heuvel,S. (2007) Replication licensing: oops!. I did it again. Curr Biol 17, R630-2.

41.  Maiorano,D., Moreau,J. and Mechali,M. (2000) XCDT1 is required for the assembly of pre-replicative complexes in Xenopus laevis. Nature 404, 622-625.

42.  Whittaker,A.J., Royzman,I. and Orr-Weaver,T.L. (2000) Drosophila double parked: a conserved, essential replication protein that colocalizes with the origin recognition complex and links DNA replication with mitosis and the down-regulation of S phase transcripts. Genes Dev 14, 1765-1776.

43.  Maiorano,D., Krasinska,L., Lutzmann,M. and Mechali,M. (2005) Recombinant Cdt1 induces rereplication of G2 nuclei in Xenopus egg extracts. Curr Biol 15, 146-153.

44.  Davidson,I.F., Li,A. and Blow,J.J. (2006) Deregulated replication licensing causes DNA fragmentation consistent with head-to-tail fork collision. Mol Cell 24, 433-443.

45.  Vaziri,C., Saxena,S., Jeon,Y., Lee,C., Murata,K., Machida,Y., Wagle,N., Hwang,D.S. and Dutta,A. (2003) A p53-dependent checkpoint pathway prevents rereplication. Mol Cell 11, 997-1008.

46.  Yoshida,K., Takisawa,H. and Kubota,Y. (2005) Intrinsic nuclear import activity of geminin is essential to prevent re-initiation of DNA replication in Xenopus eggs. Genes Cells 10, 63-73.

47.  Arentson,E., Faloon,P., Seo,J., Moon,E., Studts,J.M., Fremont,D.H. and Choi,K. (2002) Oncogenic potential of the DNA replication licensing protein CDT1. Oncogene 21, 1150-1158.

48.  Tatsumi,Y., Sugimoto,N., Yugawa,T., Narisawa-Saito,M., Kiyono,T. and Fujita,M. (2006) Deregulation of Cdt1 induces chromosomal damage without rereplication and leads to chromosomal instability. J Cell Sci 119, 3128-3140.

49.  Aggarwal,P., Lessie,M.D., Lin,D.I., Pontano,L., Gladden,A.B., Nuskey,B., Goradia,A., Wasik,M.A., Klein-Szanto,A.J., Rustgi,A.K., Bassing,C.H. and Diehl,J.A. (2007) Nuclear accumulation of cyclin D1 during S phase inhibits Cul4-dependent Cdt1 proteolysis and triggers p53-dependent DNA rereplication. Genes Dev 21, 2908-2922.

50.  Braden,W.A., Lenihan,J.M., Lan,Z., Luce,K.S., Zagorski,W., Bosco,E., Reed,M.F., Cook,J.G. and Knudsen,E.S. (2006) Distinct action of the retinoblastoma pathway on the DNA replication machinery defines specific roles for cyclin-dependent kinase complexes in prereplication complex assembly and S-phase progression. Mol Cell Biol 26, 7667-7681.