Organization of DNA Replication

Chagin, Vadim O.; Stear, Jeffrey H.; Cardoso, M. Cristina

doi:10.1101/cshperspect.a000737

Cited by 57 publications

(58 citation statements)

References 87 publications

(84 reference statements)

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“…1B). These replication patterns were similar to the ones reported previously in A549 and other cell lines 14,21,22 The typical replication patterns were more easily recognized in images of a central slice than in maximum intensity projections of the whole nucleus (Fig. 1B, compare the left and the right column).…”

Section: Replication Under Optimal Conditionssupporting

confidence: 73%

Activation of new replication foci under conditions of replication stress

et al. 2015

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Abbreviations: Cpt, camptothecin; Tpt, topotecan; EdU, 5-ethynyl-2 0 -deoxyuridine; BrdU, 5-bromo-2 0 -deoxyuridine; PCNA, proliferating cell nuclear antigen; EGFP, Enhanced Green Fluorescent Protein; gH2AX, histone H2AX phosphorylated on Ser139.DNA damage, binding of drugs to DNA or a shortage of nucleotides can decrease the rate or completely halt the progress of replication forks. Although the global rate of replication decreases, mammalian cells can respond to replication stress by activating new replication origins. We demonstrate that a moderate level of stress induced by inhibitors of topoisomerase I, commencing in early, mid or late S-phase, induces activation of new sites of replication located within or in the immediate vicinity of the original replication factories; only in early S some of these new sites are also activated at a distance greater than 300 nm. Under high stress levels very few new replication sites are activated; such sites are located within the original replication regions. There is a large variation in cellular response to stress -while in some cells the number of replication sites increases even threefold, it decreases almost twofold in other cells. Replication stress results in a loss of PCNA from replication factories and a twofold increase in nuclear volume. These observations suggest that activation of new replication origins from the pool of dormant origins within replication cluster under conditions of mild stress is generally restricted to the original replication clusters (factories) active at a time of stress initiation, while activation of distant origins and new replication factories is suppressed.

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Section: Replication Under Optimal Conditionssupporting

confidence: 73%

Activation of new replication foci under conditions of replication stress

et al. 2015

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“…Previous analyses showed that new replication factories tend to form adjacent to the ones that recently disassembled (8). Close proximity of active replication factories to high concentrations of licensed origins could contribute to recruitment of released replication factors to these adjacent regions, consistent with a "domino" model of replication factory propagation (9).…”

Section: Discussionmentioning

confidence: 58%

Multi-step Loading of Human Minichromosome Maintenance Proteins in Live Human Cells

Symeonidou

Kotsantis

Roukos

et al. 2013

Journal of Biological Chemistry

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Background: MCM2-7 loading onto chromatin licenses origins for replication. Results: MCMs exhibit transient interactions with chromatin in late mitosis, stable binding in G 1 phase and increased loading in late G 1 phase. Conclusion: Multilevel regulation of MCM2-7 loading to chromatin occurs during mitosis and preceding the G 1 /S phase transition. Significance: The dynamics of the DNA licensing system within live human cells reveal multiple control points.

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“…Early replication patterns represent actively transcribed euchromatin, whereas late replication patterns are associated with heterochromatic regions. 17 Laser microirradiation produces significant DNA damage leading to high PCNA recruitment at the irradiated sites as shown in Figure 1C. Here, we used these live cell assays for DNA replication and repair, using fluorescent tagged PCNA constructs, to directly test the cell permeability and PCNA binding ability of PIPs.…”

Section: Resultsmentioning

confidence: 99%

A novel cell permeable DNA replication and repair marker

Herce

Rajan

Lättig-Tünnemann

et al. 2014

Nucleus

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Proliferating Cell Nuclear Antigen (PCNA) is a key protein in DNA replication and repair. The dynamics of replication and repair in live cells is usually studied introducing translational fusions of PCNA. To obviate the need for transfection and bypass the problem of difficult to transfect and/or short lived cells, we have now developed a cell permeable replication and/or repair marker. The design of this marker has three essential molecular components: (1) an optimized artificial PCNA binding peptide; (2) a cell-penetrating peptide, derived from the HIV-1 Trans Activator of Transcription (TAT); (3) an in vivo cleavable linker, linking the two peptides. The resulting construct was taken up by human, hamster and mouse cells within minutes of addition to the media. Inside the cells, the cargo separated from the vector peptide and bound PCNA effectively. Both replication and repair sites could be directly labeled in live cells making it the first in vivo cell permeable peptide marker for these two fundamental cellular processes. Concurrently, we also introduced a quick peptide based PCNA staining method as an alternative to PCNA antibodies for immunofluorescence applications. In summary, we present here a versatile tool to instantaneously label repair and replication processes in fixed and live cells.

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Organization of DNA Replication

Cited by 57 publications

References 87 publications

Activation of new replication foci under conditions of replication stress

Activation of new replication foci under conditions of replication stress

Multi-step Loading of Human Minichromosome Maintenance Proteins in Live Human Cells

A novel cell permeable DNA replication and repair marker

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