The Matrix Attachment Region in the Chinese Hamster Dihydrofolate Reductase Origin of Replication May Be Required for Local Chromatid Separation

Centered in the Chinese hamster dihydrofolate reductase origin of replication is a prominent nuclear matrix attachment region (MAR). Indirect lines of evidence suggested that this MAR might be required for origin activation in early S phase. To test this possibility, we have deleted the MAR from a C...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 100; no. 6; pp. 3281 - 3286
Main Authors Mesner, L. D., Hamlin, J. L., Dijkwel, P. A.
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 18.03.2003
National Acad Sciences
The National Academy of Sciences
Subjects
Animals
Base Sequence
Binding Sites - genetics
Biological Sciences
Cell division
Cell lines
CHO Cells
Chromatids
Chromatids - genetics
Chromatids - metabolism
Chromatin
Chromosomes
Chromosomes, Artificial, Bacterial - genetics
Chromosomes, Artificial, Bacterial - metabolism
Cricetinae
DNA
DNA - genetics
DNA - metabolism
DNA Replication - genetics
Gels
Genes
Genetic loci
In Situ Hybridization, Fluorescence
Interphase
Matrix
Molecular Sequence Data
Nuclear Matrix - metabolism
Replication Origin
Rodents
S Phase
Sequence Deletion
Tetrahydrofolate Dehydrogenase - genetics
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Summary:Centered in the Chinese hamster dihydrofolate reductase origin of replication is a prominent nuclear matrix attachment region (MAR). Indirect lines of evidence suggested that this MAR might be required for origin activation in early S phase. To test this possibility, we have deleted the MAR from a Chinese hamster ovary variant harboring a single copy of the dihydrofolate reductase locus. However, 2D gel replicon mapping shows that removal of the MAR has no significant effect either on the frequency or timing of initiation in this locus. Rather, fluorescence in situ hybridization studies on cells swollen under either neutral or alkaline conditions show that deletion of the MAR interferes with local separation of daughter chromatids. This surprising result provides direct genetic evidence that at least a subset of MARs performs an important biological function, possibly related to chromatid cohesion and separation.
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To whom correspondence should be addressed. E-mail: jlh2d@virginia.edu.
Communicated by Nicholas R. Cozzarelli, University of California, Berkeley, CA
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0437791100