Journal
EXPERIMENTAL CELL RESEARCH
Volume 313, Issue 1, Pages 109-120Publisher
ELSEVIER INC
DOI: 10.1016/j.yexcr.2006.09.020
Keywords
DNA replication; origin; DHFR; dinucleotide repeat; transcription factor; ori-beta
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Funding
- NATIONAL CANCER INSTITUTE [T32CA009385] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM052948] Funding Source: NIH RePORTER
- NCI NIH HHS [T32 CA009385, T32 CA09385-20] Funding Source: Medline
- NIGMS NIH HHS [GM 52948, R01 GM052948] Funding Source: Medline
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The Chinese hamster dihydrofolate reductase (DHFR) DNA replication initiation region, the 5.8 kb ori-beta, can function as a DNA replicator at random ectopic chromosomal sites in hamster cells. We report a detailed genetic analysis of the DiNucleotide Repeat (DNR) element, one of several sequence elements necessary for ectopic ori-beta activity. Deletions within ori-beta identified a 132 bp core region within the DNR element, consisting mainly of dinucleotide repeats, and a downstream region that are required for ori-beta initiation activity at non-specific ectopic sites in hamster cells. Replacement of the DNR element with Xenopus or mouse transcriptional elements from rDNA genes restored full levels of initiation activity, but replacement with a nucleosome positioning element or a viral intron sequence did not. The requirement for the DNR element and three other ori-beta sequence elements was conserved when ori-beta activity was tested at either random sites or at a single specific ectopic chromosomal site in human cells. These results confirm the importance of specific cis-acting elements in directing the initiation of DNA replication in mammalian cells, and provide new evidence that transcriptional elements can functionally substitute for one of these elements in ori-beta. (c) 2006 Elsevier Inc. All rights reserved.
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