A site and strand specific nuclease activity with analogies to topoisomerase I frames the rRNA gene of Tetrahymena.
Exposure of macronuclear chromatin from Tetrahymena thermophila to sodium dodecyl sulfate causes an endogenous nuclease to cleave the extra-chromosomal rDNA at specific sites. All cuts are single-strand cleavages specific to the non-coding strand. Three cleavages map in the central non-transcribed spacer of the palindromic molecule at positions -1000, -600 and -150 bp with respect to the transcription initiation point. A fourth site is located close to the transcription termination point, while no cleavage is observed in the coding region. The position of each cleavage is in the immediate neighbourhood of DNAse I hypersensitive sites. Additionally, certain DNA sequence motifs are repeated in the region around the cleavages. Upon cleavage induction a protein becomes attached to the rDNA. Our results indicate covalent binding to the generated 3' end, in analogy to the aborted reaction of topoisomerase I.
D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004264:DNA Topoisomerases, Type I / Q000378:metabolism*; D004275:DNA, Ribosomal; D005796:Genes; D009693:Nucleic Acid Hybridization; D012335:RNA, Ribosomal / Q000235:genetics*; D012967:Sodium Dodecyl Sulfate; D013379:Substrate Specificity; D013768:Tetrahymena / Q000201:enzymology*; D014158:Transcription, Genetic
D004275:DNA, Ribosomal; D012335:RNA, Ribosomal; D012967:Sodium Dodecyl Sulfate; D004247:DNA; D004262:DNA Restriction Enzymes; D004264:DNA Topoisomerases, Type I