Identification of a Preinitiation Step in DNA Replication That Is Independent of Origin Recognition Complex and cdc6, but Dependent on cdk2

doi:10.1083/jcb.140.2.271

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© The Rockefeller University Press, 0021-9525/1998//271 $5.00
The Journal of Cell Biology, Volume 140, Number 2, , 1998 271-281

Article

Identification of a Preinitiation Step in DNA Replication That Is Independent of Origin Recognition Complex and cdc6, but Dependent on cdk2

Xuequn Helen Hua and John Newport

Biology Department, University of California, San Diego, CA 92093-0347

Before initiation of DNA replication, origin recognition complex(ORC) proteins, cdc6, and minichromosome maintenance (MCM) proteinsbind to chromatin sequentially and form preinitiation complexes.Using Xenopus laevis egg extracts, we find that after the formationof these complexes and before initiation of DNA replication,cdc6 is rapidly removed from chromatin, possibly degraded bya cdk2-activated, ubiquitin-dependent proteolytic pathway.If this displacement is inhibited, DNA replication fails toinitiate. We also find that after assembly of MCM proteinsinto preinitiation complexes, removal of the ORC from DNAdoes not block the subsequent initiation of replication. Importantly,under conditions in which both ORC and cdc6 protein are absentfrom preinitiation complexes, DNA replication is still dependenton cdk2 activity. Therefore, the final steps in the processleading to initiation of DNA replication during S phase ofthe cell cycle are independent of ORC and cdc6 proteins, butdependent on cdk2 activity.

Abbreviations used in this paper: ELB, egg lysis buffer; MCM, minichromosome maintenance; ORC, origin recognition complex.

Address all correspondence to J. Newport, Biology Department,0347, University of California, San Diego, 9500 Gilman Drive,La Jolla, CA 92093-0347. Tel.: (619) 534-3423. Fax: (619) 534-0555.E-mail: jnewport{at}ucsd.edu

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Remus, D., Blanchette, M., Rio, D. C., Botchan, M. R. (2005). CDK Phosphorylation Inhibits the DNA-binding and ATP-hydrolysis Activities of the Drosophila Origin Recognition Complex. J. Biol. Chem. 280: 39740-39751 [Abstract] [Full Text]
Sibani, S., Price, G. B., Zannis-Hadjopoulos, M. (2005). Decreased origin usage and initiation of DNA replication in haploinsufficient HCT116 Ku80+/- cells. J. Cell Sci. 118: 3247-3261 [Abstract] [Full Text]
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Gaczynska, M., Osmulski, P. A., Jiang, Y., Lee, J.-K., Bermudez, V., Hurwitz, J. (2004). Atomic force microscopic analysis of the binding of the Schizosaccharomyces pombe origin recognition complex and the spOrc4 protein with origin DNA. Proc. Natl. Acad. Sci. USA 101: 17952-17957 [Abstract] [Full Text]
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Francon, P., Lemaitre, J.-M., Dreyer, C., Maiorano, D., Cuvier, O., Mechali, M. (2004). A hypophosphorylated form of RPA34 is a specific component of pre-replication centers. J. Cell Sci. 117: 4909-4920 [Abstract] [Full Text]
Li, C.-j., Vassilev, A., DePamphilis, M. L. (2004). Role for Cdk1 (Cdc2)/Cyclin A in Preventing the Mammalian Origin Recognition Complex's Largest Subunit (Orc1) from Binding to Chromatin during Mitosis. Mol. Cell. Biol. 24: 5875-5886 [Abstract] [Full Text]
Sugimoto, N., Tatsumi, Y., Tsurumi, T., Matsukage, A., Kiyono, T., Nishitani, H., Fujita, M. (2004). Cdt1 Phosphorylation by Cyclin A-dependent Kinases Negatively Regulates Its Function without Affecting Geminin Binding. J. Biol. Chem. 279: 19691-19697 [Abstract] [Full Text]
Oehlmann, M., Score, A. J., Blow, J. J. (2004). The role of Cdc6 in ensuring complete genome licensing and S phase checkpoint activation. JCB 165: 181-190 [Abstract] [Full Text]
Forsburg, S. L. (2004). Eukaryotic MCM Proteins: Beyond Replication Initiation. Microbiol. Mol. Biol. Rev. 68: 109-131 [Abstract] [Full Text]
Alexandrow, M. G., Hamlin, J. L. (2004). Cdc6 Chromatin Affinity Is Unaffected by Serine-54 Phosphorylation, S-Phase Progression, and Overexpression of Cyclin A. Mol. Cell. Biol. 24: 1614-1627 [Abstract] [Full Text]
Whittaker, S. R., Walton, M. I., Garrett, M. D., Workman, P. (2004). The Cyclin-dependent Kinase Inhibitor CYC202 (R-Roscovitine) Inhibits Retinoblastoma Protein Phosphorylation, Causes Loss of Cyclin D1, and Activates the Mitogen-activated Protein Kinase Pathway. Cancer Res. 64: 262-272 [Abstract] [Full Text]
Harvey, K. J., Newport, J. (2003). Metazoan Origin Selection: ORIGIN RECOGNITION COMPLEX CHROMATIN BINDING IS REGULATED BY CDC6 RECRUITMENT AND ATP HYDROLYSIS. J. Biol. Chem. 278: 48524-48528 [Abstract] [Full Text]
Prokhorova, T. A., Mowrer, K., Gilbert, C. H., Walter, J. C. (2003). DNA replication of mitotic chromatin in Xenopus egg extracts. Proc. Natl. Acad. Sci. USA 100: 13241-13246 [Abstract] [Full Text]
Tatsumi, Y., Ohta, S., Kimura, H., Tsurimoto, T., Obuse, C. (2003). The ORC1 Cycle in Human Cells: I. CELL CYCLE-REGULATED OSCILLATION OF HUMAN ORC1. J. Biol. Chem. 278: 41528-41534 [Abstract] [Full Text]
Harvey, K. J., Newport, J. (2003). CpG Methylation of DNA Restricts Prereplication Complex Assembly in Xenopus Egg Extracts. Mol. Cell. Biol. 23: 6769-6779 [Abstract] [Full Text]
Vashee, S., Cvetic, C., Lu, W., Simancek, P., Kelly, T. J., Walter, J. C. (2003). Sequence-independent DNA binding and replication initiation by the human origin recognition complex. Genes Dev. 17: 1894-1908 [Abstract] [Full Text]
Christensen, T. W., 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 [Abstract] [Full Text]
Shimada, K., Pasero, P., Gasser, S. M. (2002). ORC and the intra-S-phase checkpoint: a threshold regulates Rad53p activation in S phase. Genes Dev. 16: 3236-3252 [Abstract] [Full Text]
Chou, D. M., Petersen, P., Walter, J. C., Walter, G. (2002). Protein Phosphatase 2A Regulates Binding of Cdc45 to the Prereplication Complex. J. Biol. Chem. 277: 40520-40527 [Abstract] [Full Text]
Edwards, M. C., Tutter, A. V., Cvetic, C., Gilbert, C. H., Prokhorova, T. A., Walter, J. C. (2002). MCM2-7 Complexes Bind Chromatin in a Distributed Pattern Surrounding the Origin Recognition Complex in Xenopus Egg Extracts. J. Biol. Chem. 277: 33049-33057 [Abstract] [Full Text]
Bell, S. P. (2002). The origin recognition complex: from simple origins to complex functions. Genes Dev. 16: 659-672 [Full Text]
Fujita, M., Ishimi, Y., Nakamura, H., Kiyono, T., Tsurumi, T. (2002). Nuclear Organization of DNA Replication Initiation Proteins in Mammalian Cells. J. Biol. Chem. 277: 10354-10361 [Abstract] [Full Text]
Jackman, M., Kubota, Y., den Elzen, N., Hagting, A., Pines, J. (2002). Cyclin A- and Cyclin E-Cdk Complexes Shuttle between the Nucleus and the Cytoplasm. Mol. Biol. Cell 13: 1030-1045 [Abstract] [Full Text]
Li, C.-J., DePamphilis, M. L. (2002). Mammalian Orc1 Protein Is Selectively Released from Chromatin and Ubiquitinated during the S-to-M Transition in the Cell Division Cycle. Mol. Cell. Biol. 22: 105-116 [Abstract] [Full Text]
Kong, D., DePamphilis, M. L. (2001). Site-Specific DNA Binding of the Schizosaccharomyces pombe Origin Recognition Complex Is Determined by the Orc4 Subunit. Mol. Cell. Biol. 21: 8095-8103 [Abstract] [Full Text]
Chaudhuri, B., Xu, H., Todorov, I., Dutta, A., Yates, J. L. (2001). Human DNA replication initiation factors, ORC and MCM, associate with oriP of Epstein-Barr virus. Proc. Natl. Acad. Sci. USA 10.1073/pnas.181347998v1 [Abstract] [Full Text]
Bryant, J. A., Moore, K., Aves, S. J. (2001). Origins and complexes: the initiation of DNA replication. J Exp Bot 52: 193-202 [Abstract] [Full Text]
Laman, H., Coverley, D., Krude, T., Laskey, R., Jones, N. (2001). Viral Cyclin-Cyclin-Dependent Kinase 6 Complexes Initiate Nuclear DNA Replication. Mol. Cell. Biol. 21: 624-635 [Abstract] [Full Text]
Lei, M, Tye, B. (2001). Initiating DNA synthesis: from recruiting to activating the MCM complex. J. Cell Sci. 114: 1447-1454 [Abstract]
Bielinsky, A., Gerbi, S. (2001). Where it all starts: eukaryotic origins of DNA replication. J. Cell Sci. 114: 643-651 [Abstract]
Snaith, H. A., Brown, G. W., Forsburg, S. L. (2000). Schizosaccharomyces pombe Hsk1p Is a Potential Cds1p Target Required for Genome Integrity. Mol. Cell. Biol. 20: 7922-7932 [Abstract] [Full Text]
Jares, P., Blow, J. J. (2000). Xenopus Cdc7 function is dependent on licensing but not on XORC, XCdc6, or CDK activity and is required for XCdc45 loading. Genes Dev. 14: 1528-1540 [Abstract] [Full Text]
Santoni-Rugiu, E., Falck, J., Mailand, N., Bartek, J., Lukas, J. (2000). Involvement of Myc Activity in a G1/S-Promoting Mechanism Parallel to the pRb/E2F Pathway. Mol. Cell. Biol. 20: 3497-3509 [Abstract] [Full Text]
Pereverzeva, I., Whitmire, E., Khan, B., Coué, M. (2000). Distinct Phosphoisoforms of the Xenopus Mcm4 Protein Regulate the Function of the Mcm Complex. Mol. Cell. Biol. 20: 3667-3676 [Abstract] [Full Text]
Zou, L., Stillman, B. (2000). Assembly of a Complex Containing Cdc45p, Replication Protein A, and Mcm2p at Replication Origins Controlled by S-Phase Cyclin-Dependent Kinases and Cdc7p-Dbf4p Kinase. Mol. Cell. Biol. 20: 3086-3096 [Abstract] [Full Text]
Lane, M. E., Elend, M., Heidmann, D., Herr, A., Marzodko, S., Herzig, A., Lehner, C. F. (2000). A Screen for Modifiers of Cyclin E Function in Drosophila melanogaster Identifies Cdk2 Mutations, Revealing the Insignificance of Putative Phosphorylation Sites in Cdk2. Genetics 155: 233-244 [Abstract] [Full Text]
Maiorano, D., Lemaitre, J.-M., Mechali, M. (2000). Stepwise Regulated Chromatin Assembly of MCM2-7 Proteins. J. Biol. Chem. 275: 8426-8431 [Abstract] [Full Text]
Romanowski, P., Marr, J., Madine, M. A., Rowles, A., Blow, J. J., Gautier, J., Laskey, R. A. (2000). Interaction of Xenopus Cdc2{middle dot}Cyclin A1 with the Origin Recognition Complex. J. Biol. Chem. 275: 4239-4243 [Abstract] [Full Text]
Prokhorova, T. A., Blow, J. J. (2000). Sequential MCM/P1 Subcomplex Assembly Is Required to Form a Heterohexamer with Replication Licensing Activity. J. Biol. Chem. 275: 2491-2498 [Abstract] [Full Text]
Gillespie, P. J., Blow, J. J. (2000). Nucleoplasmin-mediated chromatin remodelling is required for Xenopus sperm nuclei to become licensed for DNA replication. Nucleic Acids Res 28: 472-480 [Abstract] [Full Text]
Lu, Z. H., Xu, H., Leno, G. H. (1999). DNA Replication in Quiescent Cell Nuclei: Regulation by the Nuclear Envelope and Chromatin Structure. Mol. Biol. Cell 10: 4091-4106 [Abstract] [Full Text]
Ogawa, Y., Takahashi, T., Masukata, H. (1999). Association of Fission Yeast Orp1 and Mcm6 Proteins with Chromosomal Replication Origins. Mol. Cell. Biol. 19: 7228-7236 [Abstract] [Full Text]
Elsasser, S., Chi, Y., Yang, P., Campbell, J. L. (1999). Phosphorylation Controls Timing of Cdc6p Destruction: A Biochemical Analysis. Mol. Biol. Cell 10: 3263-3277 [Abstract] [Full Text]
Fujita, M., Yamada, C., Goto, H., Yokoyama, N., Kuzushima, K., Inagaki, M., Tsurumi, T. (1999). Cell Cycle Regulation of Human CDC6 Protein. INTRACELLULAR LOCALIZATION, INTERACTION WITH THE HUMAN MCM COMPLEX, AND CDC2 KINASE-MEDIATED HYPERPHOSPHORYLATION. J. Biol. Chem. 274: 25927-25932 [Abstract] [Full Text]
Pasero, P., Duncker, B. P., Schwob, E., Gasser, S. M. (1999). A role for the Cdc7 kinase regulatory subunit Dbf4p in the formation of initiation-competent origins of replication. Genes Dev. 13: 2159-2176 [Abstract] [Full Text]
Herbig, U., Marlar, C. A., Fanning, E. (1999). The Cdc6 Nucleotide-Binding Site Regulates Its Activity in DNA Replication in Human Cells. Mol. Biol. Cell 10: 2631-2645 [Abstract] [Full Text]
Snaith, H. A., Forsburg, S. L. (1999). Rereplication Phenomenon in Fission Yeast Requires MCM Proteins and Other S Phase Genes. Genetics 152: 839-851 [Abstract] [Full Text]
Leone, G., DeGregori, J., Jakoi, L., Cook, J. G., Nevins, J. R. (1999). Collaborative role of E2F transcriptional activity and G1 cyclindependent kinase activity in the induction of S phase. Proc. Natl. Acad. Sci. USA 96: 6626-6631 [Abstract] [Full Text]
Jiang, W., Wells, N. J., Hunter, T. (1999). Multistep regulation of DNA replication by Cdk phosphorylation of HsCdc6. Proc. Natl. Acad. Sci. USA 96: 6193-6198 [Abstract] [Full Text]
Chesnokov, I., Gossen, M., Remus, D., Botchan, M. (1999). Assembly of functionally active Drosophila origin recognition complex from recombinant proteins. Genes Dev. 13: 1289-1296 [Abstract] [Full Text]
Gant, T. M., Harris, C. A., Wilson, K. L. (1999). Roles of LAP2 Proteins in Nuclear Assembly and DNA Replication: Truncated LAP2{beta} Proteins Alter Lamina Assembly, Envelope Formation, Nuclear Size, and DNA Replication Efficiency in Xenopus laevis Extracts. JCB 144: 1083-1096 [Abstract] [Full Text]
Trowbridge, P. W., Roy, R., Simmons, D. T. (1999). Human Topoisomerase I Promotes Initiation of Simian Virus 40�DNA Replication In Vitro. Mol. Cell. Biol. 19: 1686-1694 [Abstract] [Full Text]
Rowles, A, Tada, S, Blow, J. (1999). Changes in association of the Xenopus origin recognition complex with chromatin on licensing of replication origins. J. Cell Sci. 112: 2011-2018 [Abstract]
Voitenleitner, C., Rehfuess, C., Hilmes, M., O'Rear, L., Liao, P.-C., Gage, D. A., Ott, R., Nasheuer, H.-P., Fanning, E. (1999). Cell Cycle-Dependent Regulation of Human DNA Polymerase alpha -Primase Activity by Phosphorylation. Mol. Cell. Biol. 19: 646-656 [Abstract] [Full Text]
Michael, W. M., Newport, J. (1998). Coupling of Mitosis to the Completion of S Phase Through Cdc34-Mediated Degradation of Wee1. Science 282: 1886-1889 [Abstract] [Full Text]
Ritzi, M., Baack, M., Musahl, C., Romanowski, P., Laskey, R. A., Knippers, R. (1998). Human Minichromosome Maintenance Proteins and Human Origin Recognition Complex 2�Protein on Chromatin. J. Biol. Chem. 273: 24543-24549 [Abstract] [Full Text]
Cueille, N., Nougarede, R., Mechali, F., Philippe, M., Bonne-Andrea, C. (1998). Functional Interaction between the Bovine Papillomavirus Virus Type 1�Replicative Helicase E1 and Cyclin E-Cdk2. J. Virol. 72: 7255-7262 [Abstract] [Full Text]
Kreitz, S., Ritzi, M., Baack, M., Knippers, R. (2001). The Human Origin Recognition Complex Protein 1 Dissociates from Chromatin during S Phase in HeLa Cells. J. Biol. Chem. 276: 6337-6342 [Abstract] [Full Text]
Ishimi, Y., Komamura-Kohno, Y. (2001). Phosphorylation of Mcm4 at Specific Sites by Cyclin-dependent Kinase Leads to Loss of Mcm4,6,7 Helicase Activity. J. Biol. Chem. 276: 34428-34433 [Abstract] [Full Text]
Chaudhuri, B., Xu, H., Todorov, I., Dutta, A., Yates, J. L. (2001). Human DNA replication initiation factors, ORC and MCM, associate with oriP of Epstein-Barr virus. Proc. Natl. Acad. Sci. USA 98: 10085-10089 [Abstract] [Full Text]
Frolova, N. S., Schek, N., Tikhmyanova, N., Coleman, T. R. (2002). Xenopus Cdc6 Performs Separate Functions in Initiating DNA Replication. Mol. Biol. Cell 13: 1298-1312 [Abstract] [Full Text]