HDAC Activity Is Required for Efficient Core Promoter Function at the Mouse Mammary Tumor Virus Promoter
HDAC inhibition by TSA impairs reinitiation of transcription from the MMTV promoter in vitro. (a) In vitro transcription was carried out with the AdML template. Nuclear extracts were mixed with the template, and PIC assembly was allowed to proceed for 1 h. Sarkosyl was added either 2 minutes before or 10 seconds after NTP addition. Transcription was allowed to proceed for 1 h. (b) In vitro transcription was carried out in the presence or absence of sarkosyl (0.05%) added after NTP addition using either the MMTV or AdML templates as described in Section 2. Transcription levels in the presence of sarkosyl are expressed relative to levels of transcription measured in the absence of sarkosyl. (c, d) Transcription of the MMTV template was carried out using extracts from either control or TSA-treated cells in the presence or absence of sarkosyl (0.05%). In (c), transcription levels are expressed relative to those measured in control extracts in the absence of sarkosyl. In (d), transcription levels measured in extracts from TSA-treated cells are expressed relative to those measured in control extracts, both in the presence of sarkosyl. The results shown were obtained from three independent experiments, and error bars represent SEM.