Real-Time monitoring of transcriptional activity in poplar transactivation assays
Using a stable reporter line, based on the circadian clock gene PtaLHY2, we developed a methodology to real time monitoring the regulation of transcription and signal transduction. With this tool, we can quickly evaluate the activity of potential regulators of diurnal and circadian rhythms in poplar.
Transient transactivation assays have been used in several plant species to evaluate transcription factor activity by analyzing a single time point. Real-time monitoring transcriptional activity in transient assays opens new perspectives for the analysis of transcriptional regulation in plants.
We created and validated a stable luciferase reporter poplar line based on the circadian clock gene
PtaLHY2, which can be used to investigate transcriptional regulation and signal transduction pathway in poplar. Using this line as a genetic background, we established a methodology to rapidly assess potential regulators of diurnal and circadian rhythms. Thus, we showed that chromatin remodeler HMGB2/3 promotes the transcriptional activation of PtaLHY2 in a gate-dependent manner.
Determination of PtaHMGB2/3 and PtaHMGB6 protein abundance followed by pPtaLHY2::LUC reporter assay