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PMID: 1461730 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Analysis of the autonomous replication behavior in human cells of the dihydrofolate reductase putative chromosomal origin of replication.

Nucleic acids research ·Vol. 20 ·No. 22 ·1992-11-25 ·Pages 5971-8

Caddle MS, Calos MP

Abstract

Chinese hamster genomic DNA sequences from the region downstream of the dihydrofolate reductase (DHFR) gene reported to contain a chromosomal origin of bidirectional DNA replication (OBR-1) were tested for their ability to support autonomous DNA replication in human cells. A 13.3 kilobase fragment containing OBR-1 and surrounding sequences supported replication in short-term and long-term replication assays, while a 4.5 kb fragment containing OBR-1 did not support substantial replication in either assay. These results are consistent with our previous observations that large fragments of human DNA support replication, while smaller fragments are less efficient. The replication activities of plasmids containing OBR-1 were no greater than those of randomly chosen human fragments of similar size. Furthermore, two-dimensional gel analysis of plasmids containing OBR-1 indicated that initiation does not preferentially occur within the OBR-1 region. These results suggest that in the context of autonomous replication, the DHFR sequences tested do not contain genetic information specifying site-specific replication initiation. Possible implications of these results for chromosomal replication are discussed.

Related Genes
DHFR
MeSH Terms
Animals Cell Line Chromosomes/metabolism Cloning, Molecular Cricetinae DNA Replication Electrophoresis, Gel, Two-Dimensional Humans Plasmids Replicon Restriction Mapping Tetrahydrofolate Dehydrogenase/genetics,metabolism
Chemicals
Tetrahydrofolate Dehydrogenase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Caddle M S
Department of Genetics, Stanford University School of Medicine, CA 94305.
Calos M P
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/20.22.5971
Published
1992-11-25
Pages
5971-8
Language
English
Region
England
NLM ID
0411011
PMCID
PMC334462
Subset
IM
Grants
NCI NIH HHS · CA 09302 · United States
External Links
PubMed source DOI: 10.1093/nar/20.22.5971 PMC Full Text View journal details
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