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PMID: 10207074 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The one-kilobase DNA fragment upstream of the ardC actin gene of Physarum polycephalum is both a replicator and a promoter.

Molecular and cellular biology ·Vol. 19 ·No. 5 ·1999-05-00 ·Pages 3506-14

Pierron G, Pallotta D, Bénard M

Abstract

The 1-kb DNA fragment upstream of the ardC actin gene of Physarum polycephalum promotes the transcription of a reporter gene either in a transient-plasmid assay or as an integrated copy in an ectopic position, defining this region as the transcriptional promoter of the ardC gene (PardC). Since we mapped an origin of replication activated at the onset of S phase within this same fragment, we examined the pattern of replication of a cassette containing the PardC promoter and the hygromycin phosphotransferase gene, hph, integrated into two different chromosomal sites. In both cases, we show by two-dimensional agarose gel electrophoresis that an efficient, early activated origin coincides with the ectopic PardC fragment. One of the integration sites was a normally late-replicating region. The presence of the ectopic origin converted this late-replicating domain into an early-replicating domain in which replication forks propagate with kinetics indistinguishable from those of the native PardC replicon. This is the first demonstration that initiation sites for DNA replication in Physarum correspond to cis-acting replicator sequences. This work also confirms the close proximity of a replication origin and a promoter, with both functions being located within the 1-kb proximal region of the ardC actin gene. A more precise location of the replication origin with respect to the transcriptional promoter must await the development of a functional autonomously replicating sequence assay in Physarum.

MeSH Terms
Actins/genetics Animals DNA Replication/genetics DNA, Protozoan/genetics Drug Resistance/genetics Kinetics Physarum polycephalum/genetics Plasmids/genetics Promoter Regions, Genetic/genetics Replication Origin/genetics S Phase/genetics Transcription, Genetic Transformation, Genetic
Chemicals
Actins DNA, Protozoan
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Pierron G
Laboratoire Organisation Fonctionnelle du Noyau, UPR-9044, CNRS, Institut de Recherches sur le Cancer, 94801 Villejuif, France. [email protected]
Pallotta D
Bénard M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.19.5.3506
Email
[email protected]
Published
1999-05-00
Pages
3506-14
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC84143
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.19.5.3506 PMC Full Text View journal details
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