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Visualization of chromatin domains created by the gypsy insulator of Drosophila

Insulators might regulate gene expression by establishing and maintaining the organization of the chromatin fiber within the nucleus. Biochemical fractionation and in situ high salt extraction of lysed cells show that two known protein components of the gypsy insulator are present in the nuclear mat...

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Autores principales: Byrd, Keith, Corces, Victor G.
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2003
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2173808/
https://www.ncbi.nlm.nih.gov/pubmed/12925706
http://dx.doi.org/10.1083/jcb.200305013
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author Byrd, Keith
Corces, Victor G.
author_facet Byrd, Keith
Corces, Victor G.
author_sort Byrd, Keith
collection PubMed
description Insulators might regulate gene expression by establishing and maintaining the organization of the chromatin fiber within the nucleus. Biochemical fractionation and in situ high salt extraction of lysed cells show that two known protein components of the gypsy insulator are present in the nuclear matrix. Using FISH with DNA probes located between two endogenous Su(Hw) binding sites, we show that the intervening DNA is arranged in a loop, with the two insulators located at the base. Mutations in insulator proteins, subjecting the cells to a brief heat shock, or destruction of the nuclear matrix lead to disruption of the loop. Insertion of an additional gypsy insulator in the center of the loop results in the formation of paired loops through the attachment of the inserted sequences to the nuclear matrix. These results suggest that the gypsy insulator might establish higher-order domains of chromatin structure and regulate nuclear organization by tethering the DNA to the nuclear matrix and creating chromatin loops.
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spelling pubmed-21738082008-05-01 Visualization of chromatin domains created by the gypsy insulator of Drosophila Byrd, Keith Corces, Victor G. J Cell Biol Article Insulators might regulate gene expression by establishing and maintaining the organization of the chromatin fiber within the nucleus. Biochemical fractionation and in situ high salt extraction of lysed cells show that two known protein components of the gypsy insulator are present in the nuclear matrix. Using FISH with DNA probes located between two endogenous Su(Hw) binding sites, we show that the intervening DNA is arranged in a loop, with the two insulators located at the base. Mutations in insulator proteins, subjecting the cells to a brief heat shock, or destruction of the nuclear matrix lead to disruption of the loop. Insertion of an additional gypsy insulator in the center of the loop results in the formation of paired loops through the attachment of the inserted sequences to the nuclear matrix. These results suggest that the gypsy insulator might establish higher-order domains of chromatin structure and regulate nuclear organization by tethering the DNA to the nuclear matrix and creating chromatin loops. The Rockefeller University Press 2003-08-18 /pmc/articles/PMC2173808/ /pubmed/12925706 http://dx.doi.org/10.1083/jcb.200305013 Text en Copyright © 2003, The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Article
Byrd, Keith
Corces, Victor G.
Visualization of chromatin domains created by the gypsy insulator of Drosophila
title Visualization of chromatin domains created by the gypsy insulator of Drosophila
title_full Visualization of chromatin domains created by the gypsy insulator of Drosophila
title_fullStr Visualization of chromatin domains created by the gypsy insulator of Drosophila
title_full_unstemmed Visualization of chromatin domains created by the gypsy insulator of Drosophila
title_short Visualization of chromatin domains created by the gypsy insulator of Drosophila
title_sort visualization of chromatin domains created by the gypsy insulator of drosophila
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2173808/
https://www.ncbi.nlm.nih.gov/pubmed/12925706
http://dx.doi.org/10.1083/jcb.200305013
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