Nucleic Acids Research, Vol 27, Issue 10 2091-2098, Copyright © 1999 by Oxford University Press
M Benard and G Pierron
It was previously shown that the two members of the cell cycle- regulated
histone H4 gene family, H4-1 and H4-2, are replicated at the onset of S
phase in the naturally synchronous plasmodium of Physarum polycephalum,
suggesting that they are flanked by replication origins. It was further
shown that a DNA fragment upstream of the H4-1 gene is able to confer
autonomous replication of a plasmid in the budding yeast. In this paper, we
re-investigated replication of the unlinked Physarum histone H4 genes by
mapping the replication origin of these two loci using alkaline agarose gel
and neutral/neutral 2-dimensional agarose gel electrophoreses. We showed
that the two replicons containing the H4 genes are simultaneously activated
at the onset of S phase and we mapped an efficient, bidirectional
replication origin in the vicinity of each gene. Our data demonstrated that
the Physarum sequence that functions as an ARS in yeast is not the site of
replication initiation at the H4-1 locus. We also observed a stalling of
the rightward moving replication fork downstream of the H4-1 gene, in a
region where transient topoisomerase II sites were previously mapped. Our
results further extend the concept of replication/transcription coupling in
Physarum to cell cycle-regulated genes.
ARTICLES
Early activated replication origins within the cell cycle-regulated histone H4 genes in Physarum
Laboratoire Organisation Fonctionnelle du Noyau, CNRS UPR-1983, IFR- 1221, F-94801 Villejuif, France. benard@infobiogen.fr
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