Physical relationship between replicons and transcription units in Physarum polycephalum.

Electron microscope spread preparations of nuclear chromatin derived from early S-phase of Physarum reveal 'beads on a string' for nonreplicated and a portion of newly replicated chromatin. Many of the early replicons contain transcription units as visualized by nascent transcripts. They are, in most cases, arranged in continuous length gradients on both newly replicated strands of chromatin, the putative origin of replication being within the transcription unit. Preferential release of DNA as acid precipitable material by DNAse I and of RNA polymerase B (estimated as release of labeled alpha-amanitin bound to isolated nuclei) is observed in early S-phase, but only if DNA synthesis is not inhibited. Also, generation of a small particle (peak A) by staphylococcal nuclease, characteristic of transcriptionally active chromatin, depends on concomitant replication of early replicons. It is concluded that DNA replication is a prerequisite for its transcription by RNA polymerase B. Thus, the sequential replication of the genome of Physarum dictates the order of transcription during S-phase which may in part control the proliferative mitotic cycle of Physarum.