Localization of ORC1 During the Cell Cycle in Human Leukemia Cells

The interaction of the origin recognition complex (ORC) with replication origins is a critical parameter in eukaryotic replication initiation. In mammals the ORC remains bound except during mitosis, thus the localization of ORC complexes allows localization of origins. A monoclonal antibody that rec...

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Main Authors: Frederick D. Coffman, Mai-Ling Reyes, Monique Brown, W. Clark Lambert, Stanley Cohen
Format: Article
Language:English
Published: Wiley 2011-01-01
Series:Analytical Cellular Pathology
Online Access:http://dx.doi.org/10.3233/ACP-2011-0041
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author Frederick D. Coffman
Mai-Ling Reyes
Monique Brown
W. Clark Lambert
Stanley Cohen
author_facet Frederick D. Coffman
Mai-Ling Reyes
Monique Brown
W. Clark Lambert
Stanley Cohen
author_sort Frederick D. Coffman
collection DOAJ
description The interaction of the origin recognition complex (ORC) with replication origins is a critical parameter in eukaryotic replication initiation. In mammals the ORC remains bound except during mitosis, thus the localization of ORC complexes allows localization of origins. A monoclonal antibody that recognizes human ORC1 was used to localize ORC complexes in populations of human MOLT-4 cells separated by cell cycle position using centrifugal elutriation. ORC1 staining in cells in early G1 is diffuse and primarily peripheral. As the cells traverse G1, ORC1 accumulates and becomes more localized towards the center of the nucleus, however around the G1/S boundary the staining pattern changes and ORC1 appears peripheral. By mid to late S phase ORC1 immunofluorescence is again concentrated at the nuclear center. During anaphase, ORC1 staining is localized mainly in the pericentriolar regions. These findings suggest that concerted movements of origin DNA sequences in addition to the previously documented assembly and disassembly of protein complexes are an important aspect of replication initiation loci in eukaryotes.
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spelling doaj-art-71d06c8c344f44b3afabb77a0faf11d12025-08-20T02:06:31ZengWileyAnalytical Cellular Pathology2210-71772210-71852011-01-0134635536110.3233/ACP-2011-0041Localization of ORC1 During the Cell Cycle in Human Leukemia CellsFrederick D. Coffman0Mai-Ling Reyes1Monique Brown2W. Clark Lambert3Stanley Cohen4Department of Pathology and Laboratory Medicine, UMDNJ - New Jersey Medical School, Newark, NJ, USAJersey City Medical Center, Jersey City, NJ, USASanofi Pasteur, Discovery Drive, Swiftwater, PA, USADepartment of Pathology and Laboratory Medicine, UMDNJ - New Jersey Medical School, Newark, NJ, USADepartment of Pathology and Laboratory Medicine, UMDNJ - New Jersey Medical School, Newark, NJ, USAThe interaction of the origin recognition complex (ORC) with replication origins is a critical parameter in eukaryotic replication initiation. In mammals the ORC remains bound except during mitosis, thus the localization of ORC complexes allows localization of origins. A monoclonal antibody that recognizes human ORC1 was used to localize ORC complexes in populations of human MOLT-4 cells separated by cell cycle position using centrifugal elutriation. ORC1 staining in cells in early G1 is diffuse and primarily peripheral. As the cells traverse G1, ORC1 accumulates and becomes more localized towards the center of the nucleus, however around the G1/S boundary the staining pattern changes and ORC1 appears peripheral. By mid to late S phase ORC1 immunofluorescence is again concentrated at the nuclear center. During anaphase, ORC1 staining is localized mainly in the pericentriolar regions. These findings suggest that concerted movements of origin DNA sequences in addition to the previously documented assembly and disassembly of protein complexes are an important aspect of replication initiation loci in eukaryotes.http://dx.doi.org/10.3233/ACP-2011-0041
spellingShingle Frederick D. Coffman
Mai-Ling Reyes
Monique Brown
W. Clark Lambert
Stanley Cohen
Localization of ORC1 During the Cell Cycle in Human Leukemia Cells
Analytical Cellular Pathology
title Localization of ORC1 During the Cell Cycle in Human Leukemia Cells
title_full Localization of ORC1 During the Cell Cycle in Human Leukemia Cells
title_fullStr Localization of ORC1 During the Cell Cycle in Human Leukemia Cells
title_full_unstemmed Localization of ORC1 During the Cell Cycle in Human Leukemia Cells
title_short Localization of ORC1 During the Cell Cycle in Human Leukemia Cells
title_sort localization of orc1 during the cell cycle in human leukemia cells
url http://dx.doi.org/10.3233/ACP-2011-0041
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