Conservation of major nuclease S1-sensitive sites in the non-conserved spacer region of ribosomal DNA in Drosophila species

Abstract

We have analysed nuclease S1-sensitive sites in cloned ribosomal DNA repeats from Drosophila melanogaster, D. hydei and D. virilis. All species contain major S1-sensitive sites in the spacer near the region of transcription termination, albeit with somewhat different positions and sensitivities. The same sites are also sensitive to the single-strand specificity of Bal31 nuclease at neutral pH. Additional major sites exist at each end of the intervening sequence within the 28 S gene of non-transcribed intervening-sequence-positive ribosomal DNA units of D. hydei. Only minor sites, however, were detected in the Pol I promoter regions. This is in contrast to Pol II transcribed genes, where S1 hypersensitivity becomes apparent at the 5' ends during gene expression. We have sequenced and mapped the S1 sites in the D. hydei spacer. They consist mainly of alternating A and T nucleotides that could form small cruciform structures. Cross-hybridization at low stringencies between the relevant S1-sensitive spacer regions of the three species indicates that the sites lie within very divergent sequences. We discuss the potential functional significance of S1 sites in rDNA spacers and intervening sequences, and the manner in which they might be maintained during rDNA sequence divergence.