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PMID: 7760799 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The Chinese hamster dihydrofolate reductase origin consists of multiple potential nascent-strand start sites.

Molecular and cellular biology ·Vol. 15 ·No. 6 ·1995-06-00 ·Pages 3023-31

Dijkwel PA, Hamlin JL

Abstract

Previous two-dimensional gel replicon-mapping studies on the amplified dihydrofolate reductase (DHFR) domain in CHOC 400 cells suggested that replication can initiate at any of a large number of sites scattered throughout a 55-kb region lying between two convergently transcribed genes. It could be argued that this unusual distributive initiation mode is unique to amplified chromosomal loci. In this paper, we report the first application of the two-dimensional gel techniques to the analysis of a single-copy locus in mammalian cells. Results obtained with both synchronized and exponentially growing CHO cells suggest that (i) initiation can also occur at any of a large number of sites distributed throughout the intergenic region in the nonamplified DHFR locus, (ii) initiation is confined to the first 2 to 2.5 h of the S period, and (iii) initiation occurs only in a fraction of the DHFR loci in each cell cycle.

MeSH Terms
Animals CHO Cells Cricetinae Cricetulus DNA/analysis Electrophoresis, Gel, Two-Dimensional Peptide Mapping Rabbits Replication Origin Tetrahydrofolate Dehydrogenase/chemistry,genetics
Chemicals
DNA Tetrahydrofolate Dehydrogenase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Dijkwel P A
Biochemistry Department, University of Virginia School of Medicine, Charlottesville 22908, USA.
Hamlin J L
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.15.6.3023
Published
1995-06-00
Pages
3023-31
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC230533
Subset
IM
Grants
NIGMS NIH HHS · GM26108 · United States
External Links
PubMed source DOI: 10.1128/MCB.15.6.3023 PMC Full Text View journal details
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