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Nucleic Acids Research, 1987, Vol. 15, No. 15 6273-6292
© 1987

Articles

S1 nuclease sensitivity of a double-stranded telomeric DNA sequence

Marcia Budarf* and Elizabeth Blackburn

Department of Molecular Biology, University of California Berkeley, CA 94720, USA

Received March 24, 1987. Revised July 10, 1987. Accepted July 10, 1987.

We examined structural properties of poly d(C4A 2).d(T 2G 4), the telomeric DNA sequence of the ciliated protozoan Tetrahymena. Under conditions of high negative supercoiling, poly d(C4A 2)*d(T2 G4) inserted in a circular plasmid vector was preferentially sensitive to digestion with S1 nuclease. Only the C4A 2 strand was sensitive to first-strand S1 cutting, with a markedly skewed pattern of hypersensitive sites in tracts of either 46 or 7 tandem repeats. Linear poly d(C4A 2)*(T2G 4) showed no preferential S1 sensitivity, no circular dichroism spectra indicative of a Z-DNA conformation, no unusual Tm, and no unusual migration in polyacrylamide gel electrophoresis. The S1 nuclease sensitivity properties are consistent with a model proposed previously for supercoiled poly d(CT).d(AG) (Pulleyblank et al., Cell 42: 271–280, 1985), consisting of a double-stranded, protonated, right-handed underwound helix. We propose that this structure is shared by related telomeric sequences and may play a role in their biological recognition.

*Present address: Department of Cytogenetics, Children's Hospital of Philadelphia, 34th and Civic Center Blvd, Philadelphia, PA 19104, USA


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