Modeling apoptotic chromatin condensation in normal cell nuclei. Requirement for intranuclear mobility and actin involvement.
Hallmarks of the terminal stages of apoptosis are genomic DNA fragmentation and chromatin condensation. Here, we have studied the mechanism of condensation both in vitro and in vivo. We found that DNA fragmentation per se of isolated nuclei from non-apoptotic cells induced chromatin condensation that closely resembles the morphology seen in apoptotic cells, independent of ATP utilization, at physiological ionic strengths. Interestingly, chromatin condensation was accompanied by release of nuclear actin, and both condensation and actin release could be blocked by reversibly pretreating nuclei with Ca2+, Cu2+, diamide, or low pH, procedures shown to stabilize internal nuclear components. Moreover, specific inhibition of nuclear F-actin depolymerization or promotion of its formation also reduced chromatin condensation. Chromatin condensation could also be inhibited by exposing nuclei to reagents that bind to the DNA minor groove, disrupting native nucleosomal DNA wrapping. In addition, in cultured cells undergoing apoptosis, drugs that inhibit depolymerization of actin or bind to the minor groove also reduced chromatin condensation, but not DNA fragmentation. Therefore, the ability of chromatin fragments with intact nucleosomes to form large clumps of condensed chromatin during apoptosis requires the apparent disassembly of internal nuclear structures that may normally constrain chromosome subdomains in non-apoptotic cells.
Widlak
Piotr
P
Department of Experimental and Clinical Radiobiology, Center of Oncology, 44-100 Gliwice, Poland.
Palyvoda
Olena
O
Kumala
Slawomir
S
Garrard
William T
WT
D000199:Actins / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D017209:Apoptosis; D002118:Calcium / Q000378:metabolism; D002412:Cations; D002467:Cell Nucleus / Q000378:metabolism* / Q000648:ultrastructure; D002843:Chromatin / Q000378:metabolism / Q000648:ultrastructure*; D003300:Copper / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D053938:DNA Fragmentation; D003958:Diamide / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D018922:HL-60 Cells; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007477:Ions; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000648:ultrastructure; D013997:Time Factors
D000199:Actins; D002412:Cations; D002843:Chromatin; D007477:Ions; D009707:Nucleosomes; D003958:Diamide; D003300:Copper; D000255:Adenosine Triphosphate; D004247:DNA; D002118:Calcium