DOI: 10.1242/jcs.113.5.887. Selective activation of pre-replication complexes in vitro at specific sites in mammalian nuclei

Selective activation of pre-replication complexes in vitro at specific sites in mammalian nuclei

10.1242/jcs.113.5.887

Crossref journal-article

The Company of Biologists

Journal of Cell Science (237)

Abstract

ABSTRACT As the first step in determining whether or not pre-replication complexes are assembled at specific sites along mammalian chromosomes, nuclei from G1-phase hamster cells were incubated briefly in Xenopus egg extract in order to initiate DNA replication. Most of the nascent DNA consisted of RNA-primed DNA chains 0.5 to 2 kb in length, and its origins in the DHFR gene region were mapped using both the early labeled fragment assay and the nascent strand abundance assay. The results revealed three important features of mammalian replication origins. First, Xenopus egg extract can selectively activate the same origins of bi-directional replication (e.g. ori-β and β′) that are used by hamster cells in vivo. Previous reports of a broad peak of nascent DNA centered at ori-β/β′ appeared to result from the use of aphidicolin to synchronize nuclei and from prolonged exposure of nuclei to egg extracts. Second, these sites were not present until late G1-phase of the cell division cycle, and their appearance did not depend on the presence of Xenopus Orc proteins. Therefore, hamster pre-replication complexes appear to be assembled at specific chromosomal sites during G1-phase. Third, selective activation of ori-β in late G1-nuclei depended on the ratio of Xenopus egg extract to nuclei, revealing that epigenetic parameters such as the ratio of initiation factors to DNA substrate could determine the number of origins activated.

Bibliography

Li, C.-J., Bogan, J. A., Natale, D. A., & DePamphilis, M. L. (2000). Selective activation of pre-replication complexes in vitro at specific sites in mammalian nuclei. Journal of Cell Science, 113(5), 887â898.

Authors 4

Cong-Jun Li (first)
Joseph A. Bogan (additional)
Darren A. Natale (additional)
Melvin L. DePamphilis (additional)

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Dates

Type	When
Created	4 years, 4 months ago (April 23, 2021, 11:19 p.m.)
Deposited	1 year, 6 months ago (Feb. 27, 2024, 7:29 a.m.)
Indexed	1 year, 5 months ago (March 5, 2024, 3:01 p.m.)
Issued	25 years, 6 months ago (March 1, 2000)
Published	25 years, 6 months ago (March 1, 2000)
Published Online	25 years, 6 months ago (March 1, 2000)
Published Print	25 years, 6 months ago (March 1, 2000)

Funders 0

None

BibTeX

@article{Li_2000, title={Selective activation of pre-replication complexes in vitro at specific sites in mammalian nuclei}, volume={113}, ISSN={1477-9137}, url={http://dx.doi.org/10.1242/jcs.113.5.887}, DOI={10.1242/jcs.113.5.887}, number={5}, journal={Journal of Cell Science}, publisher={The Company of Biologists}, author={Li, Cong-Jun and Bogan, Joseph A. and Natale, Darren A. and DePamphilis, Melvin L.}, year={2000}, month=mar, pages={887–898} }

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