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Nucleic Acids Research, 1988, Vol. 16, No. 14 7013-7024
© 1988

Articles

Analysis of the psoralen-crosslinking pattern in chromatin DNA by exonuclease digestion

R.M. Widmer, Th. Koller and J.M. Sogo

Institut für Zellbiologie, Eidgenössische Technische Hochschule, ETHZ, ETH-Hönggerberg 8093 Zürich, Switzlerland

Received February 15, 1988. Revised March 31, 1988. Accepted March 31, 1988.

When chromatin is photoreacted with psoralen, crosslinks occur preferentially in the linker DNA between nucleosomes. The pattern of these crosslinks can be analysed by exonuclease digestion of random DNA fragments, since the exonucleases tested stop at sites of psoralen-crosslinks. Further digestion of these fragments with Sl-nuclease leads to DNA fragments of nucleosomal and polynucleosomal size, which presumably carry psoralen-crosslinks at both ends. This method of analysis of chromatin structure complements the classical micrococcal nuclease digestion analysis, since it can be performed in vitro as well as in vivo, and since it is independent of pH and ionic conditions.


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