Unified bursting strategies in ectopic and endogenouseven-skippedexpression patterns

Transcription often occurs in bursts as gene promoters switch stochastically between active and inactive states. Enhancers can dictate transcriptional activity in animal development through the modulation of burst frequency, duration, or amplitude. Previous studies observed that different enhancers can achieve a wide range of transcriptional outputs through the same strategies of bursting control. For example, despite responding to different transcription factors, alleven-skippedenhancers increase transcription by upregulating burst frequency and amplitude while burst duration remains largely constant. These shared bursting strategies suggest that a unified molecular mechanism constraints how enhancers modulate transcriptional output. Alternatively, different enhancers could have converged on the same bursting control strategy because of natural selection favoring one of these particular strategies. To distinguish between these two scenarios, we compared transcriptional bursting between endogenous and ectopic gene expression patterns. Because enhancers act under different regulatory inputs in ectopic patterns, dissimilar bursting control strategies between endogenous and ectopic patterns would suggest that enhancers adapted their bursting strategies to theirtrans-regulatory environment. Here, we generated ectopiceven-skippedtranscription patterns in fruit fly embryos and discovered that bursting strategies remain consistent in endogenous and ectopiceven-skippedexpression. These results provide evidence for a unified molecular mechanism shapingeven-skippedbursting strategies and serve as a starting point to uncover the realm of strategies employed by other enhancers.