Header

UZH-Logo

Maintenance Infos

Dynamic Architecture of Eukaryotic DNA Replication Forks In Vivo, Visualized by Electron Microscopy


Zellweger, Ralph; Lopes, Massimo (2018). Dynamic Architecture of Eukaryotic DNA Replication Forks In Vivo, Visualized by Electron Microscopy. In: Muzi-Falconi, Marco; Brown, Grant W. Genome Instability : Methods and Protocols. New York: Springer, 261-294.

Abstract

The DNA replication process can be heavily perturbed by several different conditions of genotoxic stress, particularly relevant for cancer onset and therapy. The combination of psoralen crosslinking and electron microscopy has proven instrumental to reveal the fine architecture of in vivo DNA replication intermediates and to uncover their remodeling upon specific conditions of genotoxic stress. The replication structures are stabilized in vivo (by psoralen crosslinking) prior to extraction and enrichment procedures, allowing their visualization at the transmission electron microscope. This chapter outlines the procedures required to visualize and interpret in vivo replication intermediates of eukaryotic genomic DNA, and includes an improved method for enrichment of replication intermediates, compared to previously used BND-cellulose columns.

Abstract

The DNA replication process can be heavily perturbed by several different conditions of genotoxic stress, particularly relevant for cancer onset and therapy. The combination of psoralen crosslinking and electron microscopy has proven instrumental to reveal the fine architecture of in vivo DNA replication intermediates and to uncover their remodeling upon specific conditions of genotoxic stress. The replication structures are stabilized in vivo (by psoralen crosslinking) prior to extraction and enrichment procedures, allowing their visualization at the transmission electron microscope. This chapter outlines the procedures required to visualize and interpret in vivo replication intermediates of eukaryotic genomic DNA, and includes an improved method for enrichment of replication intermediates, compared to previously used BND-cellulose columns.

Statistics

Citations

Dimensions.ai Metrics
10 citations in Web of Science®
11 citations in Scopus®
2 citations in Microsoft Academic
Google Scholar™

Altmetrics

Downloads

15 downloads since deposited on 25 Oct 2017
7 downloads since 12 months
Detailed statistics

Additional indexing

Item Type:Book Section, refereed, original work
Communities & Collections:04 Faculty of Medicine > Institute of Molecular Cancer Research
07 Faculty of Science > Institute of Molecular Cancer Research
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Scopus Subject Areas:Life Sciences > Molecular Biology
Life Sciences > Genetics
Language:English
Date:17 October 2018
Deposited On:25 Oct 2017 14:13
Last Modified:22 Sep 2020 09:40
Publisher:Springer
ISSN:1064-3745
ISBN:978-1-4939-7305-7
OA Status:Green
Publisher DOI:https://doi.org/10.1007/978-1-4939-7306-4_19
PubMed ID:29043630

Download

Green Open Access

Download PDF  'Dynamic Architecture of Eukaryotic DNA Replication Forks In Vivo, Visualized by Electron Microscopy'.
Preview
Content: Accepted Version
Filetype: PDF
Size: 1MB
View at publisher
Get full-text in a library