Histone H2B phosphorylation in mammalian apoptotic cells. An association with DNA fragmentation.
Histone phosphorylation was investigated in several mammalian cells undergoing apoptosis (human HL-60 and HeLa, mouse FM3A and N18 cells, and rat thymocytes). Among the four nucleosomal core histones (H2A, H2B, H3, and H4), H2B, which is not usually phosphorylated in quiescent or growing cells, was found to be phosphorylated after treatment with various apoptotic inducers. The H2B was phosphorylated around the time when nucleosomal DNA fragmentation was initiated and, like this fragmentation, was completely blocked with Z-Asp-CH(2)-DCB, an inhibitor of ICE or ICE-like caspase. The involved single phosphopeptide of H2B proved to be phosphorylatable in vitro with a protein kinase C, and the site Ser-32 was tentatively identified. Despite typical apoptotic chromatin condensation, the H3 phosphorylation was at a low level, and the sites where phosphorylation did occur did not include any mitosis-specific phosphopeptides. Phosphorylation of H4 was increased, but the other two histone proteins (H1 and H2A) were not appreciably changed. These observations imply that 1) H2B phosphorylation occurs universally in apoptotic cells and is associated with apoptosis-specific nucleosomal DNA fragmentation, 2) chromatin condensation in apoptosis occurs by a different biochemical mechanism from those operating during mitosis or premature chromosome condensation, and 3) this unique phosphorylation of H2B is a useful biochemical hallmark of apoptotic cells.
Ajiro
K
K
Laboratory of Cell Biology, Aichi Cancer Center, Research Institute, Nagoya, 464-8681 Japan. kajiro@aichi-cc.pref.aichi.jp
D000818:Animals; D017209:Apoptosis; D061945:Caspase Inhibitors; D053938:DNA Fragmentation / Q000187:drug effects; D018922:HL-60 Cells; D017405:Herpes Simplex Virus Protein Vmw65; D006657:Histones / Q000378:metabolism*; D006801:Humans; D051379:Mice; D009707:Nucleosomes / Q000378:metabolism; D010449:Peptide Mapping; D010748:Phosphopeptides / Q000302:isolation & purification; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats
D061945:Caspase Inhibitors; D017405:Herpes Simplex Virus Protein Vmw65; D006657:Histones; D009707:Nucleosomes; D010748:Phosphopeptides