The large variability in mRNA and protein levels found from both static and dynamic measurements in single cells has been largely attributed to random periods of transcription, often occurring in bursts. The cell cycle has a pronounced global role in affecting transcriptional and translational output, but how this influences transcriptional statistics from noisy promoters is unknown and generally ignored by current stochastic models. Here we show that variable transcription from the synthetic tetO promoter in S. cerevisiae is dominated by its dependence on the cell cycle. Real-time measurements of fluorescent protein at high expression levels indicate tetO promoters increase transcription rate ∼2-fold in S/G2/M similar to constitutive genes. At low expression levels, where tetO promoters are thought to generate infrequent bursts of transcription, we observe random pulses of expression restricted to S/G2/M, which are correlated between homologous promoters present in the same cell. The analysis of static, single-cell mRNA measurements at different points along the cell cycle corroborates these findings. Our results demonstrate that highly variable mRNA distributions in yeast are not solely the result of randomly switching between periods of active and inactive gene expression, but instead largely driven by differences in transcriptional activity between G1 and S/G2/M.
There is an astonishing amount of variation in the number of mRNA and protein molecules generated from particular genes between genetically identical single cells grown in the same environment. Particularly for mRNA, the large variation seen from these “noisy” genes is consistent with the idea of transcriptional bursting where transcription occurs in random, intermittent periods of high activity. There is considerable experimental support for transcriptional bursting, and it is a primary feature of stochastic models of gene expression that account for variation. Still, it has long been recognized that variation, especially in protein levels, can occur because of global differences between genetically identical cells. We show that in budding yeast, mRNA variation is driven to a large extent by differences in the transcriptional activity of a noisy gene between different phases of the cell cycle. These differences are not because of specific cell-cycle regulation, and in some cases transcription appears restricted to certain phases, leading to pulses of mRNA production. These results raise new questions about the origins of transcriptional bursting and how the statistics of gene expression are regulated in a global way by the cell cycle.