Analyses of rRNA gene chromatin in cell cycle arrested Saccharomyces cerevisiae cells

Abstract

The existence of two chromatin structures in the rDNA locus was previously demonstrated for a large variety of organisms, ranging from yeast to human. In yeast there are about 150–200 rRNA genes organized in tandem repeats. Almost half of them are transcribed and largely depleted of nucleosomes (active/open), the other half is not transcribed and is assembled in regular arrays of nucleosomes (inactive/closed). It is proposed that RNA polymerase-I (RNAPI) transcription-elongation removes nucleosomes from closed rRNA genes (opening), and that soon after DNA replication there is deposition of nucleosomes on the open rRNA genes (closing). In G1 arrested cells, nearly all rRNA genes are depleted of nucleosomes, but most of them are not transcribed (inactive/open). In relation to the research article by Charton et al. (Mutat. Res.), the data presented here are on the hydroxyurea concentration-dependent inhibition of yeast culture growth, on cell cycle arrest before completion of genome replication, and on the opening of rRNA gene chromatin. As comparison, data are presented for yeast arrested in the G1-phase of the cell cycle by the pheromone α-factor. Keywords: Alpha-factor, Chromatin, Hydroxyurea, Psoralen crosslinking, rRNA genes, RNA polymerase I