Geminin
Binds to and inactivates Cdt1, thereby regulating licensing/pre-RC formation. Only found in metazoa. (1) (2) (3)
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Synonyms |
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Molecular weight |
• Testis specific 20kDa form (4) Xenopus 25kDa (1) , 65kDa (3)
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Biochemical properties |
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Motifs |
Xenopus • destruction box , coiled coil (1)
Mammals •Coiled coli domain dimerisation region (5) • bipartite nls (5)
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Protein interactions |
•Cdt1 (2) (3) (6) in mammals preferential binding to phosphorylated form) (7)
Xenopus • mapped cdt1 interaction site (8) Mammals • Six3/ six 6 (competes cdt1) (9) (10) • Hox and polycomb proteins (11) • Brg (12) blocks brg interaction with bHLH proteins • Cyclins e a1 and b1 (13) • interaction cdt1 affected by CoQ10 (14)
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Modifications |
Xenopus • Ub broken down via apc (1) (15)
Mammals Phosphorylation (CK2) (16)
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Structure |
• crystal structure of the coiled coil domain (17) • em structures ( tetramer -dimer of dimers) (18)
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Cellular location and expression |
Xl •Degraded in mitosis by APC. May also be regulated by post-translational modification. (1) (15) – suggested that only cdt1 bound geminin s degraded free is stable (19) •Binds to chromatin via Cdt1 at same time as cdc45 and remains bound during S phase (19) •some differences from mouse since suggested to affect cdt1 binding and displacement not just binding (19)
Mammals • reports suggest either present in g1 and s gone in M. some chromatin association in absence cdt1. (20) or absent in g1 cells, increases during s/g2 (4) – reason for differences not clear perhaps different cells or synchronisation methods. • binds to chromatin via cdt1 (21) • regulation of proteolysis (22) chicken • present all phases of cell cycle: nuclear s/g2/m, cytoplasmic g1/s border (23)
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Other comments |
• On sizing columns runs as pentamer /hexamer (24)
Xl •Destabilization of geminin in mitosis is not seen in embryonic Xenopus cell cycles (19) • deplete does not give re replication (25) (26) but get centrosome over duplication (27) , although some claim low levels re replication (8) • cdt1 prevented from interacting with geminin early in cycle by ubiquitylation on exit from metaphase (ubiquitylation does not lead to geminin proteolysis but prevents its interaction with Cdt1) (15) • needs cdt1 to bind to chromatin(26) • decrease in cdt1 levels when geminin knocked out. (28) • Stoichiometry of geminin cdt complex determines whether active or not (29) • affects binding of preRC components in vitro (30) • cleaved during apoptosis (31)
Dm • overexpression gives neural differentiation and apoptosis (32)
Mammals •Role in development (Knockout mice) (33) in neural development (34) •Geminin may stabilize Cdt1 by preventing Cdt1 ubiquitylation (7) • Negative regulation by e2f/rb (35) • Affects levels p53/p21 (36) • In human lymphomas higher geminin often associated more proliferation even though overexpression in cells cases less proliferation and apoptosis • different reported effects reduced levels: a) gives s phase block plus re replication ( re initiation) (37) (38) or b) damage pathway induced in hela cells no re replication (20) need to take out cyclin A (or cdk1 plus2) (39) . • Geminin reduction causes reduction in cdt1 levels and ub in u2os cells. (7) (40) • part of the mechanism stopping endoreduplication in mouse trophoblast development (41) • role in transcription (42) • cleaved during apoptosis by caspase 3. (43)
Plants • Arabidopsis has GEM which is unrelated to geminin but also binds and serves a similar function (44) |
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Revised by |
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Last edited |
13 July 09 |
1. McGarry,T.J. and Kirschner,M.W. (1998) Geminin, an inhibitor of DNA replication, is degraded during mitosis. Cell93, 1043-1053.
2. 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.
3. 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.
4. Eward,K.L., Obermann,E.C., Shreeram,S., Loddo,M., Fanshawe,T., Williams,C., Jung,H.I., Prevost,A.T., Blow,J.J., Stoeber,K. and Williams,G.H. (2004) DNA replication licensing in somatic and germ cells. J Cell Sci 117, 5875-5886.
5. Benjamin,J.M., Torke,S.J., Demeler,B. and McGarry,T.J. (2004) Geminin has dimerization, Cdt1-binding, and destruction domains that are required for biological activity. J Biol Chem 279, 45957-45968.
6. Ferenbach,A., Li,A., Brito-Martins,M. and Blow,J.J. (2005) Functional domains of the Xenopus replication licensing factor Cdt1. Nucleic Acids Res33, 316-324.
7. Ballabeni,A., Melixetian,M., Zamponi,R., Masiero,L., Marinoni,F. and Helin,K. (2004) Human geminin promotes pre-RC formation and DNA replication by stabilizing CDT1 in mitosis. EMBO J 23, 3122-3132.
8. Kerns,S.L., Torke,S.J., Benjamin,J.M. and McGarry,T.J. (2007) Geminin prevents rereplication during xenopus development. J Biol Chem 282, 5514-5521.
9. Del Bene,F., Tessmar-Raible,K. and Wittbrodt,J. (2004) Direct interaction of geminin and Six3 in eye development. Nature 427, 745-749.
10. Yanagi,K., Mizuno,T., Tsuyama,T., Tada,S., Iida,Y., Sugimoto,A., Eki,T., Enomoto,T. and Hanaoka,F. (2005) Caenorhabditis elegans geminin homologue participates in cell cycle regulation and germ line development. J Biol Chem280, 19689-19694.
11. Luo,L., Yang,X., Takihara,Y., Knoetgen,H. and Kessel,M. (2004) The cell-cycle regulator geminin inhibits Hox function through direct and polycomb-mediated interactions. Nature 427, 749-753.
12. Seo,S., Herr,A., Lim,J.W., Richardson,G.A., Richardson,H. and Kroll,K.L. (2005) Geminin regulates neuronal differentiation by antagonizing Brg1 activity. Genes Dev19, 1723-1734.
13. Nakuci,E., Xu,M., Pujana,M.A., Valls,J. and Elshamy,W.M. (2006) Geminin is bound to chromatin in G2/M phase to promote proper cytokinesis. Int J Biochem Cell Biol38, 1207-1220.
14. Mizushina,Y., Takeuchi,T., Takakusagi,Y., Yonezawa,Y., Mizuno,T., Yanagi,K., Imamoto,N., Sugawara,F., Sakaguchi,K., Yoshida,H. and Fujita,M. (2008) Coenzyme Q10 as a potent compound that inhibits Cdt1-geminin interaction. Biochim Biophys Acta 1780, 203-213.
15. Li,A. and Blow,J.J. (2004) Negative regulation of geminin by CDK-dependent ubiquitination controls replication licensing. Cell Cycle 3, 443-445.
16. Kulartz,M., Hiller,E., Kappes,F., Pinna,L.A. and Knippers,R. (2004) Protein kinase CK2 phosphorylates the cell cycle regulatory protein Geminin. Biochem Biophys Res Commun 315, 1011-1017.
17. Thepaut,M., Maiorano,D., Guichou,J.F., Auge,M.T., Dumas,C., Mechali,M. and Padilla,A. (2004) Crystal structure of the coiled-coil dimerization motif of geminin: structural and functional insights on DNA replication regulation. J Mol Biol 342, 275-287.
18. Okorokov,A.L., Orlova,E.V., Kingsbury,S.R., Bagneris,C., Gohlke,U., Williams,G.H. and Stoeber,K. (2004) Molecular structure of human geminin. Nat Struct Mol Biol 11, 1021-1022.
19. 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.
20. Kulartz,M. and Knippers,R. (2004) The replicative regulator protein geminin on chromatin in the HeLa cell cycle. J Biol Chem 279, 41686-41694.
21. 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.
22. Ohtsubo,M., Yasunaga,S., Ohno,Y., Tsumura,M., Okada,S., Ishikawa,N., Shirao,K., Kikuchi,A., Nishitani,H., Kobayashi,M. and Takihara,Y. (2008) Polycomb-group complex 1 acts as an E3 ubiquitin ligase for Geminin to sustain hematopoietic stem cell activity. Proc Natl Acad Sci U S A 105, 10396-10401.
23. Luo,L., Uerlings,Y., Happel,N., Asli,N.S., Knoetgen,H. and Kessel,M. (2007) Regulation of geminin functions by cell cycle-dependent nuclear-cytoplasmic shuttling. Mol Cell Biol 27, 4737-4744.
24. Madine,M. and Laskey,R. (2001) Geminin bans replication licence. Nat Cell Biol3, E49-50.
25. Yamaguchi,R. and Newport,J. (2003) A role for Ran-GTP and Crm1 in blocking re-replication. Cell 113, 115-125.
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. Tachibana,K.E., Gonzalez,M.A., Guarguaglini,G., Nigg,E.A. and Laskey,R.A. (2005) Depletion of licensing inhibitor geminin causes centrosome overduplication and mitotic defects. EMBO Rep 6, 1052-1057.
28. Mihaylov,I.S., Kondo,T., Jones,L., Ryzhikov,S., Tanaka,J., Zheng,J., Higa,L.A., Minamino,N., Cooley,L. and Zhang,H. (2002) Control of DNA replication and chromosome ploidy by geminin and cyclin A. Mol Cell Biol 22, 1868-1880.
29. 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.
30. Waga,S. and Zembutsu,A. (2006) Dynamics of DNA binding of replication initiation proteins during de novo formation of pre-replicative complexes in Xenopus egg extracts. J Biol Chem 281, 10926-10934.
31. Auziol,C., Mechali,M. and Maiorano,D. (2007) Geminin is cleaved by caspase-3 during apoptosis in Xenopus egg extracts. Biochem Biophys Res Commun 361, 276-280.
32. Quinn,L.M., Herr,A., McGarry,T.J. and Richardson,H. (2001) The Drosophila Geminin homolog: roles for Geminin in limiting DNA replication, in anaphase and in neurogenesis. Genes Dev 15, 2741-2754.
33. Hara,K., Nakayama,K.I. and Nakayama,K. (2006) Geminin is essential for the development of preimplantation mouse embryos. Genes Cells 11, 1281-1293.
34. Spella,M., Britz,O., Kotantaki,P., Lygerou,Z., Nishitani,H., Ramsay,R.G., Flordellis,C., Guillemot,F., Mantamadiotis,T. and Taraviras,S. (2007) Licensing regulators Geminin and Cdt1 identify progenitor cells of the mouse CNS in a specific phase of the cell cycle. Neuroscience 147, 373-387.
35. Markey,M., Siddiqui,H. and Knudsen,E.S. (2004) Geminin is targeted for repression by the retinoblastoma tumor suppressor pathway through intragenic E2F sites. J Biol Chem 279, 29255-29262.
36. Yoshida,K., Oyaizu,N., Dutta,A. and Inoue,I. (2004) The destruction box of human Geminin is critical for proliferation and tumor growth in human colon cancer cells. Oncogene 23, 58-70.
37. Melixetian,M., Ballabeni,A., Masiero,L., Gasparini,P., Zamponi,R., Bartek,J., Lukas,J. and Helin,K. (2004) Loss of Geminin induces rereplication in the presence of functional p53. J Cell Biol 165, 473-482.
38. Lin,J.J. and Dutta,A. (2007) ATR pathway is the primary pathway for activating G2/M checkpoint induction after re-replication. J Biol Chem 282, 30357-30362.
39. Machida,Y.J. and Dutta,A. (2007) The APC/C inhibitor, Emi1, is essential for prevention of rereplication. Genes Dev 21, 184-194.
40. Zhu,W., Chen,Y. and Dutta,A. (2004) Rereplication by depletion of geminin is seen regardless of p53 status and activates a G2/M checkpoint. Mol Cell Biol 24, 7140-7150.
41. Gonzalez,M.A., Tachibana,K.E., Adams,D.J., van der Weyden,L., Hemberger,M., Coleman,N., Bradley,A. and Laskey,R.A. (2006) Geminin is essential to prevent endoreduplication and to form pluripotent cells during mammalian development. Genes Dev 20, 1880-1884.
42. Kim,M.Y., Jeong,B.C., Lee,J.H., Kee,H.J., Kook,H., Kim,N.S., Kim,Y.H., Kim,J.K., Ahn,K.Y. and Kim,K.K. (2006) A repressor complex, AP4 transcription factor and geminin, negatively regulates expression of target genes in nonneuronal cells. Proc Natl Acad Sci U S A 103, 13074-13079.
43. Roukos,V., Iliou,M.S., Nishitani,H., Gentzel,M., Wilm,M., Taraviras,S. and Lygerou,Z. (2007) Geminin cleavage during apoptosis by caspase-3 alters its binding ability to the SWI/SNF subunit Brahma. J Biol Chem 282, 9346-9357.
44. Caro,E. and Gutierrez,C. (2007) A green GEM: intriguing analogies with animal geminin. Trends Cell Biol 17, 580-585.