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PMID: 8139554 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Transcription inhibits the replication of autonomously replicating plasmids in human cells.

Molecular and cellular biology ·Vol. 14 ·No. 4 ·1994-04-00 ·Pages 2516-24

Haase SB, Heinzel SS, Calos MP

Abstract

This study addresses the effect of transcription on replication, using a system based on autonomously replicating plasmids in human cells. We added transcriptional elements from the human cytomegalovirus promoter/enhancer and the human beta-actin promoter to autonomously replicating plasmids based on human sequences and found that the transcriptional elements inhibited plasmid replication. Furthermore, conditional inhibition of plasmid replication was demonstrated by using a tetracycline-responsive promoter. We found that replication activity of plasmids carrying this promoter was inversely correlated with promoter activity. Replication activity was partially restored on plasmids when a transcriptional termination sequence was placed directly downstream of the promoter element. Transcriptional activity of the promoters and the efficacy of the terminator sequence were confirmed by using steady-state RNA analysis. These experiments suggest that transcription inhibits DNA replication on these plasmids and that the degree of inhibition is dependent on transcription strength. The possible significance of these results for chromosomal DNA replication are discussed.

MeSH Terms
Actins/genetics Cell Line Chromosomes, Human Cytomegalovirus/genetics DNA/biosynthesis DNA Replication/drug effects DNA, Viral/biosynthesis Enhancer Elements, Genetic Genes, Immediate-Early Humans Kidney Plasmids/metabolism Promoter Regions, Genetic/drug effects Restriction Mapping Terminator Regions, Genetic Tetracycline/pharmacology Transcription, Genetic
Chemicals
Actins DNA, Viral DNA Tetracycline
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Haase S B
Department of Genetics, Stanford University School of Medicine, California 94305.
Heinzel S S
Calos M P
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34 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.14.4.2516-2524.1994
Published
1994-04-00
Pages
2516-24
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC358619
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.14.4.2516-2524.1994 PMC Full Text View journal details
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