Rapid Dynamics of Signal-Dependent Transcriptional Repression by Capicua

Abstract

Highlights d ERK signaling in the Drosophila embryo antagonizes repression in a two-step process d ERK activation leads to rapid loss of a transcriptional repressor from the DNA d Transcriptional repression is reestablished once signal is removed d Signal must persist for proper gene expression and pattern formation

From heterogeneous datasets to predictive models of embryonic development

“…1d). The power of live imaging and biochemical assays is further extended by combining them with optogenetic perturbations of the signaling cascade that culminates in ERK activation [12–14]. Here we demonstrate that datasets emerging from such varied studies (Fig.…”

“…To track endogenous nuclear Cic intensity in the middle of the embryo, flies with Cic Crispr-tagged with sfGFP (previously described in [12]) were crossed to flies containing a nuclear marker, histone-RFP. Embryos from this cross have maternally deposited Cic-sfGFP and his-RFP.…”

“…1c, 1e, and supplementary figure 1). These real-time studies reveal dynamics of molecular and cellular processes at single-cell resolution and can be coupled with biochemical assays that quantify concentration and genome-wide DNA binding patterns of Cic [12] (Fig. 1d).…”

“…We used a stochastic optimization approach to obtain an ensemble of parameter vectors that minimizes the mismatch between model predictions and datasets from measurements of Cic dynamics and its effects on transcription ([17], Materials and Methods). Data from live imaging of nuclear Cic (supplementary figure 1) and time resolved chromatin immunoprecipitation measurements of Cic binding to the regulatory DNA [12] constrained the parameters related to tier 2 of the model (i.e. phoshphorylation, nucleocytoplasmic shuttling and DNA binding).…”

Steering Molecular Activity with Optogenetics: Recent Advances and Perspectives

“…www.advanced-bio.com modality allows dynamic signaling perturbation and delineation of cellular and developmental outcomes. [50] Protein Signaling Transducer: One of the most frequent targets of optogenetic signaling manipulation is the direct control of protein signaling transducer. To date, optogenetic activation of the Ras/Raf/MEK/ERK signaling axis has been achieved at both cellular and organismal levels.…”