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PMID: 8816489 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Induction of DNA replication by transcription in the region upstream of the human c-myc gene in a model replication system.

Molecular and cellular biology ·Vol. 16 ·No. 10 ·1996-10-00 ·Pages 5754-63

Ohba R, Matsumoto K, Ishimi Y

Abstract

An important relationship between transcription and initiation of DNA replication in both eukaryotes and prokaryotes has been suggested. In an attempt to understand the molecular mechanism of this interaction, we examined whether transcription can induce DNA replication in vitro by constructing a system in which both replication and transcription were combined. Relaxed circular DNA possessing a replication initiation zone located upstream of the human c-myc gene and a T7 promoter near the P1 promoter of the gene was replicated in the presence of T7 RNA polymerase. In our model system, replication was carried out with the proteins required for simian virus 40 DNA replication. DNA synthesis, which was dependent on both T7 RNA polymerase and the replication proteins, was detected mainly in the promoter and upstream regions of the c-myc gene. Blocking RNA synthesis at the initial stage of the reaction severely reduced DNA synthesis, suggesting that RNA chain elongation is required to induce DNA synthesis. The results indicated that transcription can induce DNA replication in the upstream region of the transcribed gene, most likely by introducing negative supercoiling into the region, which results in unwinding of the DNA duplex.

MeSH Terms
Bacteriophage T7/genetics DNA Probes DNA Replication DNA, Circular/biosynthesis DNA, Superhelical/metabolism DNA-Directed RNA Polymerases/metabolism Genes, myc Humans Models, Genetic Plasmids Promoter Regions, Genetic Regulatory Sequences, Nucleic Acid Restriction Mapping Sequence Deletion Simian virus 40/genetics Transcription, Genetic Viral Proteins
Chemicals
DNA Probes DNA, Circular DNA, Superhelical Viral Proteins bacteriophage T7 RNA polymerase DNA-Directed RNA Polymerases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ohba R
Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.
Matsumoto K
Ishimi Y
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.10.5754
Published
1996-10-00
Pages
5754-63
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231576
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.16.10.5754 PMC Full Text View journal details
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