ChIP-Seq mapping of H2AK5ac, H2AK9ac, DNTTIP1, MIDEAS, and LIG4 at AsiSI-induced DNA double-strand breaks in control or MiDAC-deficient cells
To test whether MiDAC-promoted H2AK5/K9 deacetylation is implicated in DSB response, we took advantage of an improved DSB-induced via AsiSI (AiDIvA) system, in which the endonuclease restriction enzyme AsiSI is fused to the estrogen receptor (ER) hormone-binding domain and its nuclear expression in HeLa cells is controlled by doxycycline and 4-hydroxy tamoxifen (4-OHT). The precise distribution of gH2AX, H2AK5ac, H2AK9ac, FLAG-DNTTIP1, FLAG-MIDEAS, and LIG4 at AsiSI-induced DNA double-strand breaks was determined by chromatin immunoprecipitation coupled high throughput sequencing (ChIP-seq). ChIP signals from ER-AsiSI-expressing AiDIvA cells (DSB group) were normalized to ER-expressing control cells (Ctrl group). Overall design: Chromatin immunoprecipitation DNA-sequencing for H2AK5ac, H2AK9ac, DNTTIP1, MIDEAS, and LIG4 at AsiSI-induced DNA double-strand breaks in control or MiDAC-deficient Hela cells