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End‐joining of reconstituted histone H2AX‐containing chromatin in vitro by soluble nuclear proteins from human cells
Author(s) -
Siino Joseph S,
Nazarov Igor B,
Zalenskaya Irina A,
Yau Peter M,
Bradbury E.Morton,
Tomilin Nikolai V
Publication year - 2002
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(02)03176-9
Subject(s) - wortmannin , chromatin , histone , phosphorylation , microbiology and biotechnology , nucleosome , in vitro , dna , histone h2a , chemistry , biology , biochemistry , phosphatidylinositol
Non‐homologous end‐joining is an important pathway for the repair of DNA double‐strand breaks. This type of DNA break is followed by the rapid phosphorylation of Ser‐139 in the histone variant H2AX to form γ‐H2AX. Here we report efficient in vitro end‐joining of reconstituted chromatin containing nucleosomes made with either H2A or H2AX. This reaction is catalyzed by nuclear extracts from human cells and this end‐joining is not suppressed by the PI‐3 kinase inhibitor wortmannin. During the end‐joining reaction H2AX is phosphorylated at Ser‐139 as detected by immunoblot with specific antibodies and this phosphorylation is inhibited by wortmannin. Therefore, in vitro the DNA end‐joining reaction appears to be independent of H2AX phosphorylation.