Mitosis is thought to be a period of transcriptional silence due to the compact nature of mitotic chromosomes and the apparent exclusion of RNA Pol II and many transcription factors from mitotic chromatin. Yet accurate reactivation of a cell's specific gene expression program is needed to reestablish functional cell identity after mitosis. The majority of studies on protein regulation and localization during mitosis have relied extensively on antibodies and cross-linking-based approaches that are known to artifactually exclude proteins from mitotic chromatin. Here we show that RNA Pol II localization in mitosis is antibody- and fixation-dependent, and that direct assessment of transcription by pulse-labeling nascent RNA reveals global, low-level mitotic transcription. We also find a hierarchy of gene reactivation as the cells transition from mitosis to their interphase amplitude of gene expression. Resetting of gene transcription during mitotic exit is coincident with enhancer transcription. Our work thus shifts focus from assessing mitotic exit as a binary transcription switch to a more nuanced concert of transcription amplitude and enhancer usage. We suggest that understanding how gene expression patterns are conserved during mitosis rests upon deciphering how transcription is maintained by promoters.
© 2017 Palozola et al.; Published by Cold Spring Harbor Laboratory Press.