Dynamics of DNA Replication Factories in Living Cells

By H. Leonhardt, H. Rahn, P. Weinzierl, A. Sporbert, Thomas Cremer, D. Zink, C. Cardoso. Journal of Cell Biology, 2000

608 citations recorded by the source corpus.

DNA replication occurs in microscopically visible complexes at discrete sites (replication foci) in the nucleus. These foci consist of DNA associated with replication machineries, i.e., large protein complexes involved in DNA replication. To study the dynamics of these nuclear replication foci in living cells, we fused proliferating cell nuclear antigen (PCNA), a central component of the replication machinery, with the green fluorescent protein (GFP). Imaging of stable cell lines expressing low levels of GFP-PCNA showed that replication foci are heterogeneous in size and lifetime. Time-lapse studies revealed that replication foci clearly differ from nuclear speckles and coiled bodies as they neither show directional movements, nor do they seem to merge or divide. These four dimensional analyses suggested that replication factories are stably anchored in the nucleus and that changes in the pattern occur through gradual, coordinated, but asynchronous, assembly and disassembly throughout S phase.

Subfields: Biophysics, Cell Biology, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/1d0e7b1521d030542a9bb4d7759369ee2a10db93

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Cell Biology

A. Kriegstein

In Molecular Biology

Citations in the corpus

Segmentation and Classification of Cell Cycle Phases in Fluorescence Imaging (2009, unknown): “label postreplication repair of damaged DNA or immunostaining these endogenous proteins to characterize all cell cycle phases [1,2,3].”
The germinal matrices in the developing dentate gyrus are composed of neuronal progenitors at distinct differentiation stages (2013, methodology): “Proliferating cell nuclear antigen (PCNA) expression (Celis and Celis, 1985; Leonhardt et al., 2000) was used to determine proliferation of the Pax-expressing cells.”
Aurora B-mediated abscission checkpoint protects against tetraploidization. (2009, unknown): “By long-term confocal time-lapse imaging of HeLa cells stably expressing H2B-mRFP and the replication factory marker mEGFP-PCNA (Leonhardt et al., 2000), we found that cells always progressed from early to late S-phase replication foci patterns and subsequently entered mitosis, never entering a seco”
Visualizing spatiotemporal dynamics of multicellular cell-cycle progression. (2008, background): “…and the subsequent transition to G2 in live cells have been developed by fusing fluorescent proteins to proliferating cell nuclear antigen (PCNA) (Leonhardt et al., 2000; Essers et al., 2005; Kisielewska et al., 2005), DNA ligase I (Easwaran et al., 2005), or the C terminus of helicase B (GE…”
Human and bacterial oxidative demethylases repair alkylation damage in both RNA and DNA (2003, background): “In S phase, hABH2 relocates to replication foci, as shown by co-transfection with proliferating-cell nuclear antigen (PCNA), which is know”