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Transcription commonly occurs in bursts, with alternating productive (ON) andquiescent (OFF) periods, governing mRNA production rates. Yet, howtranscription is regulated through bursting dynamics remains unresolved. Inthis study, we conduct real-time measurements of endogenous transcriptionalbursting with single-mRNA sensitivity. Leveraging the diverse transcriptionalactivities in early fly embryos, we uncover stringent relationships betweenbursting parameters. Specifically, we find that the durations of ON and OFFperiods are linked. Regardless of the developmental stage or body-axisposition, gene activity levels predict the average ON and OFF periods ofindividual alleles. Lowly transcribing alleles predominantly modulate OFFdurations (burst frequency), while highly transcribing alleles primarily tuneON durations (burst size). Importantly, these relationships persist even underperturbation of cis-regulatory elements or trans-factors. This suggests a novelmechanistic constraint governing bursting dynamics rather than a modularcontrol of distinct parameters by distinct regulatory processes. Our studyprovides a foundation for future investigations into the molecular mechanismsunderpinning spatiotemporal transcriptional control.

Gene Activity as the Predictive Indicator for Transcriptional Bursting Dynamics