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Nucleic Acids Research, 1992, Vol. 20, No. 13 3309-3315
© 1992

MOLECULAR BIOLOGY

Mapping of a Physarum chromosomal origin of replication tightly linked to a developmentally-regulated profilin gene

Marianne Bénard and Gérard Pierron*

Laboratoire de Biologie et infrastructure du Noyau UPR 272-CNRS, 94801 Villejuif Cedex, France

* To whom correspondence should be addressed

Received April 27, 1992. Accepted June 1, 1992.

We compared the pattern of replication of two cell-type specific profilin genes in one developmental stage of the slime moldPhysarum polycephalum. Taking advantage of the natural synchrony of S-phase within the plasmodium, we established that the actively transcribed profilin P gene is tightly linked to a chromosomal replication origin and is replicated at the onset of S-phase. In contrast, the inactive profilin A gene is not associated with a replication origin and it is duplicated in mid S-phase. Mapping by two-dimensional gel electrophoresis defines a short DNA fragment in the proximal upstream region of the profilin P gene from which bidirectional replication is initiated. We further provide an estimate of the kinetics of elongation of the replicon and demonstrate that the 2 alleles of the profilin P gene are coordinately replicated. All these results were obtained on total DNA preparations extracted from untreated cells. They provide a strong evidence for site specific initiation of DNA replication in Physarum.


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