Nucleic Acids Research, Vol 26, Issue 10 2313-2321, Copyright © 1998 by Oxford University Press
F Toledo, B Baron, MA Fernandez, AM Lachages, V Mayau, G Buttin and M Debatisse
The nature of mammalian origins of DNA replication remains controversial
and this is primarily because two-dimensional gel replicon mapping
techniques have identified broad zones of replication initiation whereas
several other techniques, such as quantitative PCR, have disclosed more
discrete sites of initiation at the same chromosomal loci. In this report
we analyze the replication of an amplified genomic region encompassing the
3'-end of the GNAI3 gene, the entire GNAT2 gene and the intergenic region
between them in exponentially growing Chinese hamster fibroblasts. These
cells express GNAI3 but not GNAT2 . The replication pattern was first
analyzed by two- dimensional neutral-alkaline gel electrophoresis.
Surprisingly, the results revealed a small preferential zone of replication
initiation, of at most 1.7 kb, located in a limited part of the GNAI3 -
GNAT2 intergenic region. Mapping of this initiation zone was then confirmed
by quantitative PCR. The agreement between the two techniques exploited
here strengthens the hypothesis that preferred sites of replication
initiation do exist in mammalian genomes.
ARTICLES
oriGNAI3: a narrow zone of preferential replication initiation in mammalian cells identified by 2D gel and competitive PCR replicon mapping techniques
Unite de Genetique Somatique (URA CNRS 1960), Institut Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France. ftoledo@pasteur.fr
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