Localization and DNA sequence of a replication origin in the rhodopsin gene locus of Chinese hamster cells

Gale, James M.; Tobey, Robert A.; D'Anna, Joseph A.

doi:10.1016/0022-2836(92)90999-z

Cited by 77 publications

(58 citation statements)

References 77 publications

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“…While it is difficult to reconcile these results, it should be pointed out that the vast majority of studies analyzing newly synthesized DNA emanating from replication start sites (by analysis either of its distribution along the chromosome or of its polarity) have led to the identification of precise sites for initiation of DNA replication. Among the identified origins are those located within the ribosomal protein S14 gene in Chinese hamster cells (39), within the Syrian hamster CAD gene (28), and near the mouse adenosine deaminase gene (8,43), the Chinese hamster rhodopsin gene (19), the human ␤-globin gene (29), the c-myc gene (40), and the human lamin B2 gene (20). In contrast, most of the studies analyzing the structure of replication intermediates by the 2D-gel technique led to the conclusion that origins are dispersed in wide genomic areas or, at best, confined to still large preferred initiation zones (11,32,34,42).…”

Section: Discussionmentioning

confidence: 99%

High-Resolution Mapping of the Origin of DNA Replication in the Hamster Dihydrofolate Reductase Gene Domain by Competitive PCR

Pelizon

Diviacco

Falaschi

et al. 1996

Molecular and Cellular Biology

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Section: Discussionmentioning

confidence: 99%

High-Resolution Mapping of the Origin of DNA Replication in the Hamster Dihydrofolate Reductase Gene Domain by Competitive PCR

Pelizon

Diviacco

Falaschi

et al. 1996

Molecular and Cellular Biology

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“…By using this technique, bidirectional replication origin was localized in a region upstream of the mouse adenosine deaminase (ADA) gene in both single chromosomal copy and amplified extrachromosomal copies (Carroll et al, 1993). It has been also shown that replication is initiated within a 5-10-kb region near the hamster rhodopsin gene by analysis of temporal order in which neighboring segments replicate (Gale et al, 1992). On the other hand, 2-D gel electrophoresis analysis has shown that replication of the DHFR gene is initiated from a large number of locations scattered within a 55-kb zone rather than from one or two unique regions (Vaughn et al, 1990;Dijkwel and Hamlin, 1992).…”

Section: Introductionmentioning

confidence: 99%

Autonomous replication of human chromosomal DNA fragments in human cells.

Masukata

Satoh

Obuse

et al. 1993

MBoC

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We have examined whether a human chromosome has distinct segments that can replicate autonomously as extrachromosomal elements. Human 293S cells were transfected with a set of human chromosomal DNA fragments of 8-15 kilobase pairs that were cloned on an Escherichia coli plasmid vector. The transfected cells were subsequently cultured in the presence of 5-bromodeoxyuridine during two cell generations, and several plasmid clones labeled in both of the daughter DNA strands were isolated. Efficiency of replication of these clones, as determined from the ratios of heavy-heavy and one-half of heavy-light molecules to total molecules recovered from density-labeled cells, was 9.4% per cell generation on the average. Replication efficiency of control clones excluded during the selection was about 2.2% and that of the vector plasmid alone was 0.3%. A representative clone plWl replicated in a semiconservative manner only one round during the S phase of the cell cycle. It replicated extrachromosomally without integration into chromosome. The human segment of the clone was composed of several subsegments that promoted autonomous replication at different efficiencies. Our results suggest that certain specific nucleotide sequences are involved in autonomous replication of human segments.

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“…This conclusion is based on the fraction of replication forks traveling in the same direction as determined by two-dimensional (2D) neutral/alkaline gels (64, 83), the ratio of Okazaki fragments that hybridize to the two strands of a unique DNA probe (6,15,20,54,88), and the ratio of long leading nascent DNA strands from forward arms of replication forks that hybridize to the two strands of a unique DNA probe (16,46,54,57). In addition, quantitative analysis of specific DNA sequences within long nascent DNA strands reveals that most of them originate bidirectionally from a small chromosomal locus (39) and that this locus can reside within an initiation zone (97).Most OBRs are contained within as little as 0.5 kb to as much as 3 kb (references 4, 6, 26, 39, 64, 87, 88, 91, and 95 and references therein), although some OBRs lie within larger regions of 5 to 11 kb (36,46,54,83). The fact that these OBRs have been identified by independent investigators using a variety of different methods gives confidence that site-specific initiation is not an artifact of the experimental conditions used to map them.…”

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confidence: 99%

Site-Specific Initiation of DNA Replication in Xenopus Egg Extract Requires Nuclear Structure

Gilbert

Miyazawa

DePamphilis

1995

Molecular and Cellular Biology

117

184

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Previous studies have shown that Xenopus egg extract can initiate DNA replication in purified DNA molecules once the DNA is organized into a pseudonucleus. DNA replication under these conditions is independent of DNA sequence and begins at many sites distributed randomly throughout the molecules. In contrast, DNA replication in the chromosomes of cultured animal cells initiates at specific, heritable sites. Here we show that Xenopus egg extract can initiate DNA replication at specific sites in mammalian chromosomes, but only when the DNA is presented in the form of an intact nucleus. Initiation of DNA synthesis in nuclei isolated from G 1 -phase Chinese hamster ovary cells was distinguished from continuation of DNA synthesis at preformed replication forks in S-phase nuclei by a delay that preceded DNA synthesis, a dependence on soluble Xenopus egg factors, sensitivity to a protein kinase inhibitor, and complete labeling of nascent DNA chains. Initiation sites for DNA replication were mapped downstream of the amplified dihydrofolate reductase gene region by hybridizing newly replicated DNA to unique probes and by hybridizing Okazaki fragments to the two individual strands of unique probes. When G 1 -phase nuclei were prepared by methods that preserved the integrity of the nuclear membrane, Xenopus egg extract initiated replication specifically at or near the origin of bidirectional replication utilized by hamster cells (dihydrofolate reductase ori-␤). However, when nuclei were prepared by methods that altered nuclear morphology and damaged the nuclear membrane, preference for initiation at ori-␤ was significantly reduced or eliminated. Furthermore, site-specific initiation was not observed with bare DNA substrates, and Xenopus eggs or egg extracts replicated prokaryotic DNA or hamster DNA that did not contain a replication origin as efficiently as hamster DNA containing ori-␤. We conclude that initiation sites for DNA replication in mammalian cells are established prior to S phase by some component of nuclear structure and that these sites can be activated by soluble factors in Xenopus eggs.Origins of DNA replication in animal viruses and single-cell eukaryotic organisms consist of specific cis-acting sequences that function independently (autonomously replicating sequences) when transferred to the appropriate cells or cell extract (25,26,58). However, attempts to identify similar sequences in multicellular eukaryotes have resulted in a paradox: while DNA synthesis is initiated at specific, genetically determined sites in cellular chromosomes, identification of cis-acting DNA sequences that control replication has proven elusive.Mapping initiation sites for DNA replication at 17 different locations in the chromosomes of mammals and flies has revealed that DNA synthesis initiates not randomly throughout cellular chromosomes but at specific DNA sites. These sites consist of a primary initiation locus referred to as the origin of bidirectional replication (OBR) surrounded by many secondary initiation loci distribute...

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Localization and DNA sequence of a replication origin in the rhodopsin gene locus of Chinese hamster cells

Cited by 77 publications

References 77 publications

High-Resolution Mapping of the Origin of DNA Replication in the Hamster Dihydrofolate Reductase Gene Domain by Competitive PCR

High-Resolution Mapping of the Origin of DNA Replication in the Hamster Dihydrofolate Reductase Gene Domain by Competitive PCR

Autonomous replication of human chromosomal DNA fragments in human cells.

Site-Specific Initiation of DNA Replication in Xenopus Egg Extract Requires Nuclear Structure

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