Visualization of replication factories attached to a nucleoskeleton

doi:10.1016/0092-8674(93)90235-I

Cell

Volume 73, Issue 2, 23 April 1993, Pages 361-373

https://doi.org/10.1016/0092-8674(93)90235-I Get rights and content

Abstract

HeLa cells in early S phase were encapsulated in agarose microbeads, permeabilized, and incubated with biotin-11-dUTP in a “physiological” buffer. Sites of DNA synthesis were then immunolabeled. As others have found, ∼ 150 focal sites of synthesis were visible in each nucleus by light microscopy; they also contained DNA polymerase α and proliferating cell nuclear antigen. Electron microscopy of thick resinless sections from which ∼ 90% of the chromatin had been removed revealed a similar number of dense, morphologically discrete ovoid bodies strung along a nucleoskeleton. The ovoids remained morphologically and functionally intact despite the removal of most of the chromatin. After 2.5 min of incubation with biotin-11-dUTP, the incorporated analog was associated only with ovoids; after 5 min it began to spread into the adjacent chromatin, which became extensively labeled after 1 hr. This provides visual evidence for polymerization “factories” fixed to a skeleton, with replication occurring as the template moves through them.

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Cell

ArticleVisualization of replication factories attached to a nucleoskeleton

Abstract

J. Biol. Chem.

Cell

Cell

J. Mol. Biol.

J. Mol. Biol.

Cell

Exp. Cell Res.

Exp. Cell Res.

Exp. Cell Res.

Cell

Exp. Cell Res.

J. Biol. Chem.

J. Ultrastruct. Res.

Identification of a novel nuclear domain

J. Cell Biol.

Nuclear protein matrix: association with newly synthesized DNA

Science

A role for the nuclear envelope in controlling DNA replication within the cell cycle

Nature

Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites

J. Cell Biol.

Discrete nuclear domains of poly(A) RNA and their relationship to the functional organization of the nucleus

J. Cell Biol.

The nucleoskeleton: artefact, passive framework or active site?

J. Cell Sci.

Structure and function of nuclear and cytoplasmic ribonucleoprotein particles

Annu. Rev. Cell Biol.

The nonchromatin substructures of the nucleus: the ribonucleoprotein (RNP)-containing and RNP-depleted matrices analyzed by sequential fractionation and resinless section microscopy

J. Cell Biol.

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Clin. Exp. Immunol.

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Article
Visualization of replication factories attached to a nucleoskeleton