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Nucleic Acids Research, 1983, Vol. 11, No. 22 7661-7678
© 1983

ENZYMOLOGY

A site and strand spedfic nuciease activity with analogies to topoisomerase I frames the rRNA gene of Tetrahymena

Elmar Gocke, Bjarne J. Bonven and Ole Westergaard

Department of Molecular Biology, University of Aarhus 8000 Aarhus C, Denmark

Received October 21, 1983. Accepted October 31, 1983.

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.


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