| Influence of the live cell DNA marker DRAQ5 on chromatin-associated processes. | |
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MedLine Citation:
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PMID: 20439168 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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In the last decade, live cell fluorescence microscopy experiments have revolutionized cellular and molecular biology, enabling the localization of proteins within cellular compartments to be analysed and to determine kinetic parameters of enzymatic reactions in living nuclei to be measured. Recently, in vivo DNA labelling by DNA-stains such as DRAQ5, has provided the opportunity to measure kinetic reactions of GFP-fused proteins in targeted areas of the nucleus with different chromatin compaction levels. To verify the suitability of combining DRAQ5-staining with protein dynamic measurements, we have tested the cellular consequences of DRAQ5 DNA intercalation. We show that DRAQ5 intercalation rapidly modifies both the localization and the mobility properties of several DNA-binding proteins such as histones, DNA repair, replication and transcription factors, by stimulating a release of these proteins from their substrate. Most importantly, the effect of DRAQ5 on the mobility of essential cellular enzymes results in a potent inhibition of the corresponding cellular functions. From these observations, we suggest that great caution must be used when interpreting live cell data obtained using DRAQ5. |
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Authors:
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Pierre-Olivier Mari; Vincent Verbiest; Simone Sabbioneda; Audrey M Gourdin; Nils Wijgers; Christoffel Dinant; Alan R Lehmann; Wim Vermeulen; Giuseppina Giglia-Mari |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-05-02 |
Journal Detail:
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Title: DNA repair Volume: 9 ISSN: 1568-7856 ISO Abbreviation: DNA Repair (Amst.) Publication Date: 2010 Jul |
Date Detail:
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Created Date: 2010-06-28 Completed Date: 2010-09-16 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101139138 Medline TA: DNA Repair (Amst) Country: Netherlands |
Other Details:
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Languages: eng Pagination: 848-55 Citation Subset: IM |
Copyright Information:
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2010 Elsevier B.V. All rights reserved. |
Affiliation:
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Department of Genetics, Erasmus MC, GE Rotterdam, The Netherlands. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Anthraquinones
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chemistry,
pharmacology* Cell Line Chromatin / chemistry*, metabolism DNA / biosynthesis, chemistry*, genetics DNA Repair / drug effects* DNA Replication / drug effects DNA-Binding Proteins / chemistry*, metabolism Histones / chemistry, metabolism Humans Intercalating Agents / chemistry*, pharmacology Staining and Labeling / methods Stress, Physiological Transcription, Genetic / drug effects |
| Grant Support | |
ID/Acronym/Agency:
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//Medical Research Council |
| Chemical | |
Reg. No./Substance:
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0/1,5-bis((2-(methylamino)ethyl)amino)-4,8-dihydroxyanthracene-9,10-dione; 0/Anthraquinones; 0/Chromatin; 0/DNA-Binding Proteins; 0/Histones; 0/Intercalating Agents; 9007-49-2/DNA |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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