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RPAP3 interacts with Reptin to regulate UV‐induced phosphorylation of H2AX and DNA damage
Author(s) -
Ni Lin,
Saeki Makio,
Xu Li,
Nakahara Hirokazu,
Saijo Masafumi,
Tanaka Kiyoji,
Kamisaki Yoshinori
Publication year - 2009
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.22073
Subject(s) - dna damage , phosphorylation , microbiology and biotechnology , cycloheximide , hela , chromatin remodeling , g2 m dna damage checkpoint , apoptosis , biology , immunoprecipitation , chromatin , hek 293 cells , dna , cell cycle checkpoint , cell cycle , cell , biochemistry , protein biosynthesis , gene
We have previously reported that Monad, a novel WD40 repeat protein, potentiates apoptosis induced by tumor necrosis factor‐α and cycloheximide. By affinity purification and mass spectrometry, RNA polymerase II‐associated protein 3 (RPAP3) was identified as a Monad binding protein and may function with Monad as a novel modulator of apoptosis pathways. Here we report that Reptin, a highly conserved AAA + ATPase that is part of various chromatin‐remodeling complexes, is also involved in the association of RPAP3 by immunoprecipitation and confocal microscopic analysis. Overexpression of RPAP3 induced HEK293 cells to death after UV‐irradiation. Loss of RPAP3 by RNAi improved HeLa cell survival after UV‐induced DNA damage and attenuated the phosphorylation of H2AX. Depletion of Reptin reduced cell survival and facilitated the phosphorylation on H2AX. These results suggest that RPAP3 modulates UV‐induced DNA damage by regulating H2AX phosphorylation. J. Cell. Biochem. 106: 920–928, 2009. © 2009 Wiley‐Liss, Inc.