Shadow enhancers modulate distinct transcriptional parameters that differentially effect downstream patterning events

• Published in: Development (Cambridge) Vol. 149; no. 21
• Main Authors: Whitney, Peter H, Shrestha, Bikhyat, Xiong, Jiahan, Zhang, Tom, Rushlow, Christine A
• Format: Journal Article
• Language: English
• Published: England The Company of Biologists Ltd 01.11.2022
• Subjects: Animals; Drosophila - metabolism; Drosophila Proteins - genetics; Drosophila Proteins - metabolism; Enhancer Elements, Genetic - genetics; Gastrulation; Gene Expression Regulation, Developmental; MS2 live imaging; Morphogen gradient; Shadow enhancers; Drosophila; Transcriptional kinetics
• ISSN: 0950-1991; 1477-9129
• DOI: 10.1242/dev.200940

Transcription in the early Drosophila blastoderm is coordinated by the collective action of hundreds of enhancers. Many genes are controlled by so-called 'shadow enhancers', which provide resilience to environment or genetic insult, allowing the embryo to robustly generate a precise transcriptional pattern. Emerging evidence suggests that many shadow enhancer pairs do not drive identical expression patterns, but the biological significance of this remains unclear. In this study, we characterize the shadow enhancer pair controlling the gene short gastrulation (sog). We removed either the intronic proximal enhancer or the upstream distal enhancer and monitored sog transcriptional kinetics. Notably, each enhancer differs in sog spatial expression, timing of activation and RNA Polymerase II loading rates. In addition, modeling of individual enhancer activities demonstrates that these enhancers integrate activation and repression signals differently. Whereas activation is due to the sum of the two enhancer activities, repression appears to depend on synergistic effects between enhancers. Finally, we examined the downstream signaling consequences resulting from the loss of either enhancer, and found changes in tissue patterning that can be explained by the differences in transcriptional kinetics measured.