Engagement of DNA polymerases during apoptosis.
DNA replicative and repair machinery was investigated by means of different techniques, including in vitro nuclear enzymatic assays, immunoelectron microscopy and confocal microscopy, in apoptotic cell lines such as HL-60 treated with methotrexate, P815 and K562 exposed to low temperatures and Friend cells exposed to ionizing radiation. The results showed a shift of DNA polymerase alpha and beta activities. DNA polymerase alpha, which in controls was found to be the principal replicative enzyme driving DNA synthesis, underwent, upon apoptosis, a large decrease of its activity being replaced by DNA polymerase beta which is believed to be associated with DNA repair. Such a modulation was concomitant with a topographical redistribution of both DNA polymerase alpha and the incorporation of BrdUrd throughout the nucleus. Taken together, these results indicate the occurrence of a dramatic response of the DNA machinery, through a possible common or at least similar behaviour when different cell lines are triggered to apoptosis. Although this possibility requires further investigation, these findings suggest an extreme attempt of the cell undergoing apoptosis to preserve its nuclear environment by switching on a repair/defence mechanism during fragmentation and chromatin margination.
Miscia
S
S
Istituto di Morfologia Umana Normale, Università G. d'Annunzio, Chieti, Italy. s.miscia@morpho.unich.it
Di Baldassarre
A
A
Alba Rana
R
R
Di Pietro
R
R
Cataldi
A
A
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000528:radiation effects; D004256:DNA Polymerase I / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D019951:DNA Polymerase beta / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D004259:DNA-Directed DNA Polymerase / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D005622:Friend murine leukemia virus; D018922:HL-60 Cells / Q000201:enzymology / Q000648:ultrastructure; D006801:Humans; D007035:Hypothermia / Q000201:enzymology / Q000473:pathology; D004915:Leukemia, Erythroblastic, Acute / Q000201:enzymology / Q000473:pathology; D012515:Mast-Cell Sarcoma / Q000648:ultrastructure; D008727:Methotrexate / Q000494:pharmacology; D051379:Mice; D014407:Tumor Cells, Cultured
D004256:DNA Polymerase I; D019951:DNA Polymerase beta; D004259:DNA-Directed DNA Polymerase; D008727:Methotrexate