Initiation of DNA fragmentation during apoptosis induces phosphorylation of H2AX histone at serine 139.
Histone H2AX is a ubiquitous member of the H2A histone family that differs from the other H2A histones by the presence of an evolutionarily conserved C-terminal motif, -KKATQASQEY. The serine residue in this motif becomes rapidly phosphorylated in cells and animals when DNA double-stranded breaks are introduced into their chromatin by various physical and chemical means. In the present communication we show that this phosphorylated form of H2AX, referred to as gamma-H2AX, appears during apoptosis concurrently with the initial appearance of high molecular weight DNA fragments. gamma-H2AX forms before the appearance of internucleosomal DNA fragments and the externalization of phosphatidylserine to the outer membrane leaflet. gamma-H2AX formation is inhibited by N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone and the inhibitor of caspase-activated DNase, and it is induced when DNase I and restriction enzymes are introduced into cells, suggesting that any apoptotic endonuclease is sufficient to induce gamma-H2AX formation. These results indicate that gamma-H2AX formation is an early chromatin modification following initiation of DNA fragmentation during apoptosis.
Rogakou
E P
EP
Laboratory of Molecular Pharmacology, Division of Basic Sciences, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA. emrog@helix.nih.gov
Nieves-Neira
W
W
Boon
C
C
Pommier
Y
Y
Bonner
W M
WM
D017209:Apoptosis; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D053938:DNA Fragmentation; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D020287:In Situ Nick-End Labeling; D010766:Phosphorylation; D012694:Serine / Q000378:metabolism*; D014162:Transfection
D006657:Histones; D012694:Serine; D020169:Caspases