Real-Time monitoring of transcriptional activity in poplar transactivation assays

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.

Original Paper:

Ramos-Sánchez, JM; Triozzi, PM; Moreno-Cortés, A; Conde, D; Perales, M; Allona, I. 2017. "Real-time monitoring of PtaHMGB activity in poplar transactivation assays". Plant Methods. DOI: 10.1186/s13007-017-0199-x".

Image: Determination of PtaHMGB2/3 and PtaHMGB6 protein abundance followed by pPtaLHY2::LUC reporter assay