Phosphorylation of histones triggers DNA fragmentation in thymocyte undergoing apoptosis induced by protein phosphatase inhibitors.
The treatment of thymocytes with protein phosphatase inhibitors such as calyculin A and okadaic acid resulted in apoptosis with a concomitant increase in phosphorylation of nuclear proteins. The phosphorylated protein in the thymocyte nuclei induced by protein phosphatase inhibitors was identified as histones by the use of two-dimensional polyacrylamide gel electrophoresis. These compounds accelerated the phosphorylation of histone H2A, H3, and H1. On the other hand, little phosphorylation of H2B and H4 by these compounds was observed. The effect of these compounds on the level of nuclear histones was also examined using high-performance capillary electrophoresis. No significant changes in the level of histones were seen in the nuclei of thymocytes treated with calyculin A and okadaic acid. Thus, the induction of thymocyte apoptosis is involved in the chemical modification of histones but not the change in their quantity. Moreover, the treatment of thymocytes with calyculin A increased the sensitivity toward endogenous DNase in the nuclei. These results suggest that phosphorylation of histones, especially H2A, H3, and H1, is an early step of triggering DNA fragmentation in thymocyte apoptosis.
Enomoto
R
R
Department of Pharmacology, High Technology Research Center, Kobe Gakuin University, Ikawadani-cho, Nishi-ku, Kobe, 651-2180, Japan.
Koyamazaki
R
R
Maruta
Y
Y
Tanaka
M
M
Takuma
K
K
Mori
K
K
Lee
E
E
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D053938:DNA Fragmentation / Q000187:drug effects*; D003851:Deoxyribonucleases / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D008387:Marine Toxins; D019319:Okadaic Acid / Q000494:pharmacology; D010080:Oxazoles / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D013950:Thymus Gland / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*
D004791:Enzyme Inhibitors; D006657:Histones; D008387:Marine Toxins; D010080:Oxazoles; D019319:Okadaic Acid; C059041:calyculin A; D003851:Deoxyribonucleases; D010749:Phosphoprotein Phosphatases