The telomeric region is excluded from nucleosomal fragmentation during apoptosis, but the bulk nuclear chromatin is randomly degraded.
Characteristic steps during cellular apoptosis are the induction of chromatin condensation and subsequent DNA fragmentation, finally leading to the formation of oligomers of nucleosomes. We have examined the kinetics and local distribution of this nucleosomal fragmentation within different genomic regions. For the induction of apoptosis, HL60 cells were treated with the water-soluble camptothecin derivative topotecan (a topoisomerase I inhibitor). The genomic origin of the fragments was analysed by Southern blot hybridisation of the cleaved DNA. In these experiments we observed similar hybridisation patterns of the fragmented DNA, indicating a random and synchronous cleavage of the nuclear chromatin. However, hybridisation with a telomeric probe revealed that, in contrast to the other analysed genomic regions, the telomeric chromatin was not cleaved into nucleosomal fragments despite our observation that the telomeric DNA in HL60 cells is organised in nucleosomes. We determined just a minor shortening of the telomeric repeats early during apoptosis. These observations suggest that telomeric chromatin is excluded from internucleosomal cleavage during apoptosis.
Schliephacke
T
T
Institut für Biochemie und Molekulare Zellbiologie, Universität Göttingen, Humboldtallee 23, D-37073 Göttingen, Germany.
Meinl
A
A
Kratzmeier
M
M
Doenecke
D
D
Albig
W
W
D016590:Aphidicolin / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018922:HL-60 Cells; D006801:Humans; D007700:Kinetics; D008836:Micrococcal Nuclease / Q000494:pharmacology; D009707:Nucleosomes / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D016615:Telomere; D019772:Topotecan / Q000494:pharmacology
D002843:Chromatin; D004791:Enzyme Inhibitors; D009707:Nucleosomes; D011993:Recombinant Fusion Proteins; D016590:Aphidicolin; D019772:Topotecan; D008836:Micrococcal Nuclease