Mcm2, but Not Rpa, Is a Component of the Mammalian Early G1-Phase Prereplication Complex

doi:10.1083/jcb.146.4.709

This Article

	Full Text
	Full Text (PDF, 780K)
	PPT slides of all figures
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JCB
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Dimitrova, D. S.
	Articles by Gilbert, D. M.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Dimitrova, D. S.
	Articles by Gilbert, D. M.


Related Collections

	Related Article

Social Bookmarking

	What's this?

© The Rockefeller University Press, 0021-9525/1999//709 $5.00
The Journal of Cell Biology, Volume 146, Number 4, , 1999 709-722

Original Article

Mcm2, but Not Rpa, Is a Component of the Mammalian Early G1-Phase Prereplication Complex

Daniela S. Dimitrova^a, Ivan T. Todorov^b, Thomas Melendy^c, and David M. Gilbert^a

^a Department of Biochemistry and Molecular Biology, S.U.N.Y. Health Science Center, Syracuse, New York 13210
^b Desmos, Inc., San Diego, California 92121
^c Department of Microbiology, S.U.N.Y. at Buffalo, School of Medicine and Biomedical Sciences, Buffalo, New York 14214
Department of Biochemistry and Molecular Biology, S.U.N.Y. Health Science Center, 750 East Adams Street, Syracuse, NY 13210.(315) 464-8750(315) 464-8723

gilbertd{at}vax.cs.hscsyr.edu

Previous experiments in Xenopus egg extracts identified whatappeared to be two independently assembled prereplication complexes(pre-RCs) for DNA replication: the stepwise assembly of ORC,Cdc6, and Mcm onto chromatin, and the FFA-1–mediated recruitmentof RPA into foci on chromatin. We have investigated whetherboth of these pre-RCs can be detected in Chinese hamster ovary(CHO) cells. Early- and late-replicating chromosomal domainswere pulse-labeled with halogenated nucleotides and prelabeledcells were synchronized at various times during the followingG1-phase. The recruitment of Mcm2 and RPA to these domains wasexamined in relation to the formation of a nuclear envelope,specification of the dihydrofolate reductase (DHFR) replicationorigin and entry into S-phase. Mcm2 was loaded gradually andcumulatively onto both early- and late-replicating chromatinfrom late telophase throughout G1-phase. During S-phase, detectableMcm2 was rapidly excluded from PCNA-containing active replicationforks. By contrast, detergent-resistant RPA foci were undetectableuntil the onset of S-phase, when RPA joined only the earliest-firingreplicons. During S-phase, RPA was present with PCNA specificallyat active replication forks. Together, our data are consistentwith a role for Mcm proteins, but not RPA, in the formationof mammalian pre-RCs during early G1-phase.

Key Words: pre-RC • Mcm • RPA • cell cycle • DNA replication domains

1.used in this paper: BrdU, bromodeoxyuridine; CldU, 5-chloro-2'-deoxyuridine;DHFR, dihydrofolate reductase; FFA, focus forming activity;IdU, 5-iodo-2'-deoxyuridine; ODP, origin decision point; ORC,origin recognition complex; PCNA, proliferating cell nuclearantigen; pre-RCs, prereplication complexes or centers; RPA,replication protein A; SSB, single-stranded DNA-binding protein;TxRed, Texas Red; xlRPA, Xenopus laevis RPA homologue

CiteULike

Complore

Connotea

Del.icio.us Add to Digg

Digg

Facebook

Technorati

Twitter What's this?

Related Article

In Brief
J. Cell Biol. 1999 146: 1-2. [Full Text] [PDF]

This article has been cited by other articles:

Lau, E., Chiang, G. G., Abraham, R. T., Jiang, W. (2009). Divergent S Phase Checkpoint Activation Arising from Prereplicative Complex Deficiency Controls Cell Survival. Mol. Biol. Cell 20: 3953-3964 [Abstract] [Full Text]
Burgess, R. C., Lisby, M., Altmannova, V., Krejci, L., Sung, P., Rothstein, R. (2009). Localization of recombination proteins and Srs2 reveals anti-recombinase function in vivo. JCB 185: 969-981 [Abstract] [Full Text]
Kanter, D. M., Bruck, I., Kaplan, D. L. (2008). Mcm Subunits Can Assemble into Two Different Active Unwinding Complexes. J. Biol. Chem. 283: 31172-31182 [Abstract] [Full Text]
Ramos-Nino, M. E., Blumen, S. R., Sabo-Attwood, T., Pass, H., Carbone, M., Testa, J. R., Altomare, D. A., Mossman, B. T. (2008). HGF Mediates Cell Proliferation of Human Mesothelioma Cells through a PI3K/MEK5/Fra-1 Pathway. Am. J. Respir. Cell Mol. Bio. 38: 209-217 [Abstract] [Full Text]
Ge, X. Q., Jackson, D. A., Blow, J. J. (2007). Dormant origins licensed by excess Mcm2 7 are required for human cells to survive replicative stress. Genes Dev. 21: 3331-3341 [Abstract] [Full Text]
Manthey, K. C., Opiyo, S., Glanzer, J. G., Dimitrova, D., Elliott, J., Oakley, G. G. (2007). NBS1 mediates ATR-dependent RPA hyperphosphorylation following replication-fork stall and collapse. J. Cell Sci. 120: 4221-4229 [Abstract] [Full Text]
Zellner, E., Herrmann, T., Schulz, C., Grummt, F. (2007). Site-specific interaction of the murine pre-replicative complex with origin DNA: assembly and disassembly during cell cycle transit and differentiation. Nucleic Acids Res 35: 6701-6713 [Abstract] [Full Text]
Barry, E. R., Bell, S. D. (2006). DNA Replication in the Archaea. Microbiol. Mol. Biol. Rev. 70: 876-887 [Abstract] [Full Text]
Maeshima, K., Yahata, K., Sasaki, Y., Nakatomi, R., Tachibana, T., Hashikawa, T., Imamoto, F., Imamoto, N. (2006). Cell-cycle-dependent dynamics of nuclear pores: pore-free islands and lamins. J. Cell Sci. 119: 4442-4451 [Abstract] [Full Text]
Braden, W. A., Lenihan, J. M., Lan, Z., Luce, K. S., Zagorski, W., Bosco, E., Reed, M. F., Cook, J. G., 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 [Abstract] [Full Text]
Tsuji, T., Ficarro, S. B., Jiang, W. (2006). Essential Role of Phosphorylation of MCM2 by Cdc7/Dbf4 in the Initiation of DNA Replication in Mammalian Cells. Mol. Biol. Cell 17: 4459-4472 [Abstract] [Full Text]
Moyer, S. E., Lewis, P. W., Botchan, M. R. (2006). Isolation of the Cdc45/Mcm2-7/GINS (CMG) complex, a candidate for the eukaryotic DNA replication fork helicase. Proc. Natl. Acad. Sci. USA 103: 10236-10241 [Abstract] [Full Text]
Dimitrova, D. S. (2006). Nuclear transcription is essential for specification of mammalian replication origins.. GENES CELLS 11: 829-844 [Abstract] [Full Text]
Woodward, A. M., Gohler, T., Luciani, M. G., Oehlmann, M., Ge, X., Gartner, A., Jackson, D. A., Blow, J. J. (2006). Excess Mcm2-7 license dormant origins of replication that can be used under conditions of replicative stress. JCB 173: 673-683 [Abstract] [Full Text]
Waga, S., 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 [Abstract] [Full Text]
Laskey, R. (2005). The Croonian Lecture 2001 Hunting the antisocial cancer cell: MCM proteins and their exploitation. Phil Trans R Soc B 360: 1119-1132 [Abstract] [Full Text]
Wilkinson, D. E., Weller, S. K. (2005). Inhibition of the Herpes Simplex Virus Type 1 DNA Polymerase Induces Hyperphosphorylation of Replication Protein A and Its Accumulation at S-Phase-Specific Sites of DNA Damage during Infection. J. Virol. 79: 7162-7171 [Abstract] [Full Text]
Naderi, S., Wang, J. Y.J., Chen, T.-T., Gutzkow, K. B., Blomhoff, H. K. (2005). cAMP-mediated Inhibition of DNA Replication and S Phase Progression: Involvement of Rb, p21Cip1, and PCNA. Mol. Biol. Cell 16: 1527-1542 [Abstract] [Full Text]
Kemp, M. G., Ghosh, M., Liu, G., Leffak, M. (2005). The histone deacetylase inhibitor trichostatin A alters the pattern of DNA replication origin activity in human cells. Nucleic Acids Res 33: 325-336 [Abstract] [Full Text]
Shechter, D., Ying, C. Y., Gautier, J. (2004). DNA Unwinding Is an MCM Complex-dependent and ATP Hydrolysis-dependent Process. J. Biol. Chem. 279: 45586-45593 [Abstract] [Full Text]
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]
Angus, S. P., Mayhew, C. N., Solomon, D. A., Braden, W. A., Markey, M. P., Okuno, Y., Cardoso, M. C., Gilbert, D. M., Knudsen, E. S. (2004). RB Reversibly Inhibits DNA Replication via Two Temporally Distinct Mechanisms. Mol. Cell. Biol. 24: 5404-5420 [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]
Shin, J.-H., Jiang, Y., Grabowski, B., Hurwitz, J., Kelman, Z. (2003). Substrate Requirements for Duplex DNA Translocation by the Eukaryal and Archaeal Minichromosome Maintenance Helicases. J. Biol. Chem. 278: 49053-49062 [Abstract] [Full Text]
Kaplan, D. L., Davey, M. J., O'Donnell, M. (2003). Mcm4,6,7 Uses a "Pump in Ring" Mechanism to Unwind DNA by Steric Exclusion and Actively Translocate along a Duplex. J. Biol. Chem. 278: 49171-49182 [Abstract] [Full Text]
Sears, J., Kolman, J., Wahl, G. M., Aiyar, A. (2003). Metaphase Chromosome Tethering Is Necessary for the DNA Synthesis and Maintenance of oriP Plasmids but Is Insufficient for Transcription Activation by Epstein-Barr Nuclear Antigen 1. J. Virol. 77: 11767-11780 [Abstract] [Full Text]
Biard, D. S.F., Miccoli, L., Despras, E., Harper, F., Pichard, E., Creminon, C., Angulo, J. F. (2003). Participation of kin17 Protein in Replication Factories and in Other DNA Transactions Mediated by High Molecular Weight Nuclear Complexes. Mol Cancer Res 1: 519-531 [Abstract] [Full Text]
Ito, S., Yanagi, K. (2003). Epstein-Barr Virus (EBV) Nuclear Antigen 1 Colocalizes with Cellular Replication Foci in the Absence of EBV Plasmids. J. Virol. 77: 3824-3831 [Abstract] [Full Text]
Liu, J.-S., Kuo, S.-R., Beerman, T. A., Melendy, T. (2003). Induction of DNA Damage Responses by Adozelesin Is S Phase-specific and Dependent on Active Replication Forks. Molecular Cancer Therapeutics 2: 41-47 [Abstract] [Full Text]
Angus, S. P., Wheeler, L. J., Ranmal, S. A., Zhang, X., Markey, M. P., Mathews, C. K., Knudsen, E. S. (2002). Retinoblastoma Tumor Suppressor Targets dNTP Metabolism to Regulate DNA Replication. J. Biol. Chem. 277: 44376-44384 [Abstract] [Full Text]
Dimitrova, D. S., Berezney, R. (2002). The spatio-temporal organization of DNA replication sites is identical in primary, immortalized and transformed mammalian cells. J. Cell Sci. 115: 4037-4051 [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]
Daniely, Y., Dimitrova, D. D., Borowiec, J. A. (2002). Stress-Dependent Nucleolin Mobilization Mediated by p53-Nucleolin Complex Formation. Mol. Cell. Biol. 22: 6014-6022 [Abstract] [Full Text]
Fortunato, E. A., Sanchez, V., Yen, J. Y., Spector, D. H. (2002). Infection of Cells with Human Cytomegalovirus during S Phase Results in a Blockade to Immediate-Early Gene Expression That Can Be Overcome by Inhibition of the Proteasome. J. Virol. 76: 5369-5379 [Abstract] [Full Text]
Keller, C., Hyrien, O., Knippers, R., Krude, T. (2002). Site-specific and temporally controlled initiation of DNA replication in a human cell-free system. Nucleic Acids Res 30: 2114-2123 [Abstract] [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]
Schwed, G., May, N., Pechersky, Y., Calvi, B. R. (2002). Drosophila Minichromosome Maintenance 6 Is Required for Chorion Gene Amplification and Genomic Replication. Mol. Biol. Cell 13: 607-620 [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]
Dimitrova, D. S., Prokhorova, T. A., Blow, J. J., Todorov, I. T., Gilbert, D. M. (2002). Mammalian nuclei become licensed for DNA replication during late telophase. J. Cell Sci. 115: 51-59 [Abstract] [Full Text]
Djeliova, V., Russev, G., Anachkova, B. (2001). Dynamics of association of origins of DNA replication with the nuclear matrix during the cell cycle. Nucleic Acids Res 29: 3181-3187 [Abstract] [Full Text]
Montecucco, A., Rossi, R., Ferrari, G., Scovassi, A. I., Prosperi, E., Biamonti, G. (2001). Etoposide Induces the Dispersal of DNA Ligase I from Replication Factories. Mol. Biol. Cell 12: 2109-2118 [Abstract] [Full Text]
Sever-Chroneos, Z., Angus, S. P., Fribourg, A. F., Wan, H., Todorov, I., Knudsen, K. E., Knudsen, E. S. (2001). Retinoblastoma Tumor Suppressor Protein Signals through Inhibition of Cyclin-Dependent Kinase 2 Activity To Disrupt PCNA Function in S Phase. Mol. Cell. Biol. 21: 4032-4045 [Abstract] [Full Text]
Dehde, S., Rohaly, G., Schub, O., Nasheuer, H.-P., Bohn, W., Chemnitz, J., Deppert, W., Dornreiter, I. (2001). Two Immunologically Distinct Human DNA Polymerase {alpha}-Primase Subpopulations Are Involved in Cellular DNA Replication. Mol. Cell. Biol. 21: 2581-2593 [Abstract] [Full Text]
Chen, C.-Y., Graham, J., Yan, H. (2001). Evidence for a Replication Function of Ffa-1, the Xenopus Orthologue of Werner Syndrome Protein. JCB 152: 985-996 [Abstract] [Full Text]
Thykjaer, T., Workman, C., Kruhøffer, M., Demtröder, K., Wolf, H., Andersen, L. D., Frederiksen, C. M., Knudsen, S., Ørntoft, T. F. (2001). Identification of Gene Expression Patterns in Superficial and Invasive Human Bladder Cancer. Cancer Res. 61: 2492-2499 [Abstract] [Full Text]
Mirzoeva, O. K., Petrini, J. H. J. (2001). DNA Damage-Dependent Nuclear Dynamics of the Mre11 Complex. Mol. Cell. Biol. 21: 281-288 [Abstract] [Full Text]
Izumi, M., Vaughan, O. A., Hutchison, C. J., Gilbert, D. M. (2000). Head and/or CaaX Domain Deletions of Lamin Proteins Disrupt Preformed Lamin A and C But Not Lamin B Structure in Mammalian Cells. Mol. Biol. Cell 11: 4323-4337 [Abstract] [Full Text]
Mendez, J., Stillman, B. (2000). Chromatin Association of Human Origin Recognition Complex, Cdc6, and Minichromosome Maintenance Proteins during the Cell Cycle: Assembly of Prereplication Complexes in Late Mitosis. Mol. Cell. Biol. 20: 8602-8612 [Abstract] [Full Text]
Loo, Y.-M., Melendy, T. (2000). The majority of human replication protein A remains complexed throughout the cell cycle. Nucleic Acids Res 28: 3354-3360 [Abstract] [Full Text]
Daniely, Y., Borowiec, J. A. (2000). Formation of a Complex between Nucleolin and Replication Protein a after Cell Stress Prevents Initiation of DNA Replication. JCB 149: 799-810 [Abstract] [Full Text]
Wang, Y., Guan, J., Wang, H., Wang, Y., Leeper, D., Iliakis, G. (2001). Regulation of DNA Replication after Heat Shock by Replication Protein A-Nucleolin Interactions. J. Biol. Chem. 276: 20579-20588 [Abstract] [Full Text]