AF9 is known to interact with multiple proteins including activators and repressors of transcription. Our data indicate that other AF9 binding proteins compete with the histone methyltransferase DOT1L for AF9 binding thus diminishing its ability to methylate lysine 79 of histone 3. Specifically, we show that AF9 is part of a protein multimer containing members of Polycomb group (PcG) PRC1 complex, CBX8, RING1B, and BMI1. Interaction with CBX8 precludes AF9-DOT1L binding. Knockdown of CBX8 with short-hairpin RNA (shRNA) leads to decreased expression of the AF9 target gene ENaCα. In contrast, CBX8 overexpression results in increased ENaCα mRNA levels and this effect can be partially overcome by co-overexpression of AF9.
Keywords: CTD; ChIP; ENaCα; Gene regulation; H3K79; Histone methylation; PRC1; PcG; PcG repressive complex 1; Polycomb-group protein; carboxy-terminal domain; chromatin immunoprecipitation; histone 3 lysine 79; polycomb group; qRT-PCR; quantitative reverse transcriptase polymerase chain reaction; shRNA; short-hairpin RNA.
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