Nucleosome positioning in the rat protamine 1 gene in vivo and in vitro

The positions of the nucleosomes along the rat protamine 1 gene have been determined in vivo through micrococcal nuclease digestion of isolated nuclei followed by Southern analysis and indirect end labeling with a protamine 1 gene probe. Several strong positioning signals are detected in rat liver nuclei where the gene is repressed. In vitro reconstitution of nucleosomes along the cloned rat protamine 1 gene results in a precisely positioned nucleosome with a dyad axis at -109 bp upstream from the transcriptional start site. The position of this nucleosome reconstituted in vitro coincides with the position of one of the nucleosomes present in vivo in rat liver nuclei. Two important regulatory elements of the expression of the protamine 1 gene, the serum response element (CArG box) and the protamine 1 consensus (cAMP response element), are positioned over the -109 nucleosome with potential functional implications for transcriptional activation.