Genomewide identification of LHY binding targets

The plant circadian clock enables plants to adjust their physiology and modulate their developmental responses in anticipation of diurnal and seasonal changes in environmental conditions. Thus, it makes an important contribution to overall fitness. The complex regulatory network underlying the oscillatory mechanism of the clock is now well characterized, but we still know relatively little about how downstream processes are controlled. In order to investigate this question we identified genes that are under direct control of one of the oscillator components, a MYB transcription factor known as LATE ELONGATED HYPOCOTYL (LHY). Overall design: The Columbia (Col-0) samples were extracted at 2 hours after dawn, corresponding to the time of the day at which LHY is maximally expressed. Chromatin immunoprecipitation experiments were carried out using an antibody to the full-length LHY protein. Input DNA was used as control in ChIP experiments.

植物昼夜节律钟能够使植物预判环境条件的昼夜与季节变化，进而调整自身生理状态并调控发育响应，因此对植物整体适合度具有重要贡献。目前，该钟振荡机制背后的复杂调控网络已得到充分解析，但我们对下游过程的调控机制仍知之甚少。为探究这一问题，我们鉴定出受昼夜节律钟核心组分之一——MYB转录因子LATE ELONGATED HYPOCOTYL（LHY）直接调控的基因。实验设计：我们在黎明后2小时（即LHY表达量达到峰值的每日时段）采集哥伦比亚（Col-0）生态型样本。本研究使用针对全长LHY蛋白的抗体开展染色质免疫共沉淀（Chromatin Immunoprecipitation, ChIP）实验，以Input DNA作为ChIP实验的对照。