Using average transcription level to understand the regulation of stochastic gene activation

Abstract

Gene activation is a random process, modelled as a framework of multiple rate-limiting steps listed sequentially, in parallel or in combination. Together with suitably assumed processes of gene inactivation, transcript birth and death, the step numbers and parameters in activation frameworks can be estimated by fitting single-cell transcription data. However, current algorithms require computing master equations that are tightly correlated with prior hypothetical frameworks of gene activation. We found that prior estimation of the framework can be facilitated by the traditional dynamical data of mRNA average level M ( t ), presenting discriminated dynamical features. Rigorous theory regarding M ( t ) profiles allows to confidently rule out the frameworks that fail to capture M ( t ) features and to test potential competent frameworks by fitting M ( t ) data. We implemented this procedure for a large number of mouse fibroblast genes under tumour necrosis factor induction and determined exactly the ‘cross-talking n -state’ framework; the cross-talk between the signalling and basal pathways is crucial to trigger the first peak of M ( t ), while the following damped gentle M ( t ) oscillation is regulated by the multi-step basal pathway. This framework can be used to fit sophisticated single-cell data and may facilitate a more accurate understanding of stochastic activation of mouse fibroblast genes.