Structural and functional preservation of specific sequences of DNA and mRNA in apoptotic bodies from ES cells.
Retinoic acid-induced apoptosis of embryonic stem (ES) cells is an experimental system which resembles the physiological programmed cell death that occurs during differentiation in embryonic development. Our aim was to analyze the involvement of epigenetic modifications such as DNA methylation and chromatin structure in the apoptotic process and to investigate the metabolic activity of apoptotic bodies. We found a relationship between DNA methylation and apoptosis, shown by a dose-dependent induction of apoptosis after treatment with the inhibitor of DNA methylation 5-aza-2'-deoxycytidine. Interestingly, we found a slight demethylation of specific sequences of the U2afl-rs1 imprinted gene in those RA treated cells which were specifically undergoing apoptosis. In addition, apoptotic bodies exhibited an unexpected open chromatin conformation accessible to the endonuclease DNase-I. Furthermore, we observed a structural and functional preservation of specific DNA sequences and mRNA. These results suggest that biological activities, such as transcription or protein synthesis, could be maintained even towards the end of the apoptotic process.
Andollo
N
N
Department of Cell Biology and Histology, Faculty of Medicine and Dentistry, University of the Basque Country, Leioa, E-48940, Vizcaya, Spain.
Boyano
M D
MD
Andrade
R
R
Zalduendo
M M
MM
Eguizabal
C
C
Asumendi
A
A
Arlucea
J
J
Aréchaga
J
J
D000818:Animals; D017209:Apoptosis; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D019921:Biotinylation; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D002454:Cell Differentiation; D049109:Cell Proliferation; D002843:Chromatin / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D019175:DNA Methylation; D000077209:Decitabine; D003850:Deoxyribonuclease I / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian / Q000166:cytology*; D005434:Flow Cytometry; D051379:Mice; D008854:Microscopy, Electron; D008957:Models, Genetic; D011419:Propidium / Q000494:pharmacology; D014176:Protein Biosynthesis; D011487:Protein Conformation; D012333:RNA, Messenger / Q000737:chemistry*; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors; D014212:Tretinoin / Q000737:chemistry
D002843:Chromatin; D012333:RNA, Messenger; D011419:Propidium; D014212:Tretinoin; D000077209:Decitabine; D004247:DNA; D003850:Deoxyribonuclease I; D001374:Azacitidine