Scaffold attachment of DNA loops in metaphase chromosomes

Details

Serval ID
serval:BIB_3C16F94535C1
Type
Article: article from journal or magazin.
Collection
Publications
Title
Scaffold attachment of DNA loops in metaphase chromosomes
Journal
Journal of Molecular Biology
Author(s)
Mirkovitch  J., Gasser  S. M., Laemmli  U. K.
ISSN
0022-2836 (Print)
Publication state
Published
Issued date
03/1988
Volume
200
Number
1
Pages
101-9
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Mar 5
Abstract
We have examined the higher-order loop organization of DNA in interphase nuclei and metaphase chromosomes from Drosophila Kc cells, and we detect no changes in the distribution of scaffold-attached regions (SARs) between these two phases of the cell cycle. The SARs, previously defined from experiments with interphase nuclei, not only are bound to the metaphase scaffold when endogenous DNA is probed but also rebind specifically to metaphase scaffolds when added exogenously as cloned, end-labeled fragments. Since metaphase scaffolds have a simpler protein pattern than interphase nuclear scaffolds, and both have a similar binding capacity, it appears that the population of proteins required for the specific scaffold-DNA interaction is limited to those found in metaphase scaffolds. Surprisingly, metaphase scaffolds isolated from Drosophila Kc cells contain both the lamin protein and a pore-complex protein, glycoprotein (gp) 188. To study whether lamin contributes to the SAR-scaffold interaction, we have carried out comparative binding studies with scaffolds from HeLa metaphase chromosomes, which are free of lamina, and from HeLa interphase nuclei. All Drosophila SAR fragments tested bind with excellent specificity to HeLa interphase scaffolds, whereas a subset of them bind to HeLa metaphase scaffolds. The maintenance of the scaffold-DNA interaction in metaphase indicates that lamin proteins are not involved in the attachment site for at least a subset of Drosophila SARs. This evolutionary and cell-cycle conservation of scaffold binding sites is consistent with a fundamental role for these fragments in the organization of the genome into looped domains.
Keywords
Animals Chromosomes/*metabolism DNA/*metabolism Drosophila melanogaster Hela Cells Humans Lamins Metaphase Nuclear Proteins/*metabolism *Nucleic Acid Conformation
Pubmed
Web of science
Create date
25/01/2008 8:41
Last modification date
20/08/2019 13:32
Usage data