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  Probing DNA structure and function with a multi-wavelength fluorescence confocal laser microscope.

Arndt-Jovin, D. J., Robertnicoud, M., & Jovin, T. M. (1990). Probing DNA structure and function with a multi-wavelength fluorescence confocal laser microscope. Journal of Microscopy-Oxford, 157, 61-72. doi:10.1111/j.1365-2818.1990.tb02947.x.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0013-0E7D-7 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-384C-0
Genre: Journal Article

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Arndt-Jovin, D. J.1, Author              
Robertnicoud, M.2, Author
Jovin, T. M.1, Author              
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1Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              
2Max Planck Society, ou_persistent13              

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 Abstract: Three levels of organization in DNA structure in the interphase cell nucleus are assessed by confocal laser scanning microscopy: (i) the conformational state of the double helix; (ii) the distribution of eu- and heterochromatin; and (iii) the localization of replication complexes throughout S phase. Multi-parameter measurements were carried out in each optical section using two laser sources and combined stereoscopic reconstructions were used to assess the co-localization of nuclear components. DNA is highly polymorphic and can adopt a variety of different helical conformations as well as unusual structures (curved, cruciform, multi-stranded). We have assessed by laser scanning microscopy the presence of left-handed Z-DNA in polytene chromosomes of Diptera as well as the spatio-temporal distribution of Z-DNA binding proteins in whole-mount Drosophila embryos and ovaries. We have determined the 3-D distribution of replication sites relative to heterochromatin regions, nucleoli and nuclear membrane by using short pulses of BrdU incorporation in synchronized mouse and human fibroblasts. Replication sites were visualized with a monoclonal anti-BrdU antibody combined with DNA fluorescent staining and antibody labelling of nuclear lamin. The implications of dynamic DNA movement and structural rearrangement to the organization of the nucleus in domains are discussed.

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Language(s): eng - English
 Dates: 1990-01
 Publication Status: Published in print
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 Table of Contents: -
 Rev. Method: Peer
 Degree: -

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Title: Journal of Microscopy-Oxford
Source Genre: Journal
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Pages: - Volume / Issue: 157 Sequence Number: - Start / End Page: 61 - 72 Identifier: ISSN: 0022-2720