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Nucleic Acids Research, 1993, Vol. 21, No. 18 4206-4209
© 1993

MOLECULAR BIOLOGY

DNA fragmentation during apoptosis is caused by frequent single-strand cuts

Manuel C. Peitsh, Christian Muüller and Jü Tschopp*

Institute of Biochemistry, University of Lausanne Chemi des Boveresses 155, CH-1066 Epalinges, Switzerland

*To whom correspondence should be addressed

Received June 16, 1993. Revised August 4, 1993. Accepted August 4, 1993.

One of the hallmarks of apoptosis Is the digestion of genomlc DNA by an endonuclease, generating a ladder of small fragments of double-stranded DNA. We have examined the nature of the DNA breaks produced In mouse thymocytes triggered to undergo apoptosis by steroids or by stimulation of the T cell receptor. Whereas the typical ladder pattern of ollgonucleosomal fragments was observed after agarose gel electrophoresis, numerous single-strand cuts were detected after electrophoresls under denaturing conditions. Single-strand nicks were found to be very frequent In the internucleosomal regions, but also to occur in the core particle-associated DNA. An Identical pattern of single-strand nicks was obtained when chromatin DNA was exposed to the single-strand cleaving deoxyribonuclease I. The nicked DNA fragments, extracted from apoptotic thymocytes, were sensitive to the action of S1nuclease. We propose that DNA fragmentation Induced during apoptosis is not due to a double-strand cutting enzyme as previously postulated, but rather is the result of single-strand breaks. This ensures the dissociation of the DNA molecule at sites where cuts are found within close proximity.


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