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PMID: 7969119 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Hairpin structures are the primary amplification products: a novel mechanism for generation of inverted repeats during gene amplification.

Molecular and cellular biology ·Vol. 14 ·No. 12 ·1994-12-00 ·Pages 7782-91

Cohen S, Hassin D, Karby S, Lavi S

Abstract

Early events of DNA amplification which occur during perturbed replication were studied by using simian virus 40 (SV40)-transformed Chinese hamster cells (CO60) as a model system. The amplification is observed shortly after carcinogen treatment, and the amplified sequences contain molecules organized as inverted repeats (IRs). SV40 amplification in vitro was studied by using extracts from carcinogen-treated CO60 cells. In the amplified DNA the SV40 origin region was rereplicated, while more distal sequences were not replicated even once. Using several experimental procedures such as sucrose gradients, "snap-back" assay, and two-dimensional gel electrophoresis, we show that the overreplicated DNA contains IRs which are synthesized de novo as hairpins or stem-loop structures which were detached from the template molecules. The fully replicated SV40 molecules synthesized by the HeLa extracts do not contain such IRs. We propose "U-turn replication" as a novel mechanism for gene amplification, accounting for the generation of extrachromosomal inverted duplications as a result of perturbed replication and template switching of the DNA polymerases.

MeSH Terms
Animals Cricetinae Cricetulus DNA Replication DNA, Viral/ultrastructure Gene Amplification HeLa Cells Humans Hydrogen Bonding In Vitro Techniques Methylnitronitrosoguanidine/pharmacology Models, Biological Nucleic Acid Conformation Simian virus 40/genetics Templates, Genetic
Chemicals
DNA, Viral Methylnitronitrosoguanidine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Cohen S
Department of Cell Research and Immunology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Ramat Aviv, Israel.
Hassin D
Karby S
Lavi S
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.14.12.7782-7791.1994
Published
1994-12-00
Pages
7782-91
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC359318
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.14.12.7782-7791.1994 PMC Full Text View journal details
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