Sequence-specific cleavage of chromatin by staphylococcal nuclease can generate an atypical nucleosome pattern.
We have investigated the nucleosomal organization of ribosomal genes in the acellular slime mold Physarum polycephalum. When probed with staphylococcal nuclease, the ribosomal genes appear to be uniformly packed in nucleosomes, in an arrangement which is indistinguishable from the pattern obtained with bulk chromatin. During this study, an unusual pattern of digestion was obtained from a DNA region immediately upstream of the initiation site of rRNA transcription, in addition to the nucleosomal profile, a second regular ladder of fragments with a repeat length of 30-40 basepairs was generated from this region. We established that this pattern of degradation reflects the strong preference of staphylococcal nuclease for certain nucleotide arrangements on the DNA, rather than a particular chromatin configuration. These observations clearly show that great caution needs to be exerted whenever data from staphylococcal nuclease digestions are interpreted in terms of chromatin structure.
D001483:Base Sequence; D002843:Chromatin / Q000032:analysis*; D005796:Genes; D008836:Micrococcal Nuclease / Q000032:analysis*; D009693:Nucleic Acid Hybridization; D009707:Nucleosomes / Q000648:ultrastructure*; D010804:Physarum / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics; D012270:Ribosomes / Q000648:ultrastructure; D014158:Transcription, Genetic
D002843:Chromatin; D009707:Nucleosomes; D012335:RNA, Ribosomal; D008836:Micrococcal Nuclease