Chromosome architecture can dictate site-specific initiation of DNA replication in Xenopus egg extracts

doi:10.1083/jcb.135.5.1207

This Article

	Full Text (PDF, 2103K)
	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 Lawlis, S. J.
	Articles by Gilbert, D. M.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Lawlis, S. J.
	Articles by Gilbert, D. M.


Social Bookmarking

	What's this?

ARTICLES

Chromosome architecture can dictate site-specific initiation of DNA replication in Xenopus egg extracts

SJ Lawlis, SM Keezer, JR Wu and DM Gilbert
Department of Biochemistry and Molecular Biology, State University of New York Health Science Center, Syracuse 13210, USA.

Xenopus egg extracts initiate DNA replication specifically at the dihydrofolate reductase (DHFR) origin locus with intact nuclei from late G1-phase CHO cells as a substrate, but at nonspecific sites when purified DNA is assembled by the extract into an embryonic nuclear structure. Here we show that late G1-phase CHO nuclei can be cycled through an in vitro Xenopus egg mitosis, resulting in the assembly of an embryonic nuclear envelope around G1-phase chromatin. Surprisingly, replication within these chimeric nuclei initiated at a novel specific site in the 5' region of the DHFR structural gene that does not function as an origin in cultured CHO cells. Preferential initiation at this unusual site required topoisomerase II-mediated chromosome condensation during mitosis. Nuclear envelope breakdown and reassembly in the absence of chromosome condensation resulted in nonspecific initiation. Introduction of condensed chromosomes from metaphase- arrested CHO cells directly into Xenopus egg extracts was sufficient to elicit assembly of chimeric nuclei and preferential initiation at this same site. These results demonstrate clearly that chromosome architecture can determine the sites of initiation of replication in Xenopus egg extracts, supporting the hypothesis that patterns of initiation in vertebrate cells are established by higher order features of chromosome structure.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

Dimitrova, D. S. (2006). Nuclear transcription is essential for specification of mammalian replication origins.. GENES CELLS 11: 829-844 [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]
Liu, G., Malott, M., Leffak, M. (2003). Multiple Functional Elements Comprise a Mammalian Chromosomal Replicator. Mol. Cell. Biol. 23: 1832-1842 [Abstract] [Full Text]
Kamath, S., Leffak, M. (2001). Multiple sites of replication initiation in the human {beta}-globin gene locus. Nucleic Acids Res 29: 809-817 [Abstract] [Full Text]
Cimbora, D. M., Schübeler, D., Reik, A., Hamilton, J., Francastel, C., Epner, E. M., Groudine, M. (2000). Long-Distance Control of Origin Choice and Replication Timing in the Human beta -Globin Locus Are Independent of the Locus Control Region. Mol. Cell. Biol. 20: 5581-5591 [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]
Ortega, J., DePamphilis, M. (1999). Nucleoskeleton and initiation of DNA replication in metazoan cells. J. Cell Sci. 111: 3663-3673 [Abstract]
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]
Vernis, L., Chasles, M., Pasero, P., Lepingle, A., Gaillardin, C., Fournier, P. (1999). Short DNA Fragments without Sequence Similarity Are Initiation Sites for Replication in the Chromosome of the Yeast Yarrowia lipolytica. Mol. Biol. Cell 10: 757-769 [Abstract] [Full Text]
Sasaki, T., Sawado, T., Yamaguchi, M., Shinomiya, T. (1999). Specification of Regions of DNA Replication Initiation during Embryogenesis in the 65-Kilobase DNApolalpha -dE2F Locus of Drosophila melanogaster. Mol. Cell. Biol. 19: 547-555 [Abstract] [Full Text]
Kobayashi, T., Rein, T., DePamphilis, M. L. (1998). Identification of Primary Initiation Sites for DNA Replication in the Hamster Dihydrofolate Reductase Gene Initiation Zone. Mol. Cell. Biol. 18: 3266-3277 [Abstract] [Full Text]
Lu, Z. H., Sittman, D. B., Romanowski, P., Leno, G. H. (1998). Histone H1 Reduces the Frequency of Initiation in Xenopus Egg Extract by Limiting the Assembly of Prereplication Complexes on Sperm Chromatin. Mol. Biol. Cell 9: 1163-1176 [Abstract] [Full Text]