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Davari, Kathrin; Lichti, Johannes; Gallus, Christian; Greulich, Franziska; Uhlenhaut, N. Henriette; Heinig, Matthias; Friedel, Caroline C.; Glasmacher, Elke
Cell reports, 04/2017, Letnik: 19, Številka: 3Journal Article
Activation of immune cells results in rapid functional changes, but how such fast changes are accomplished remains enigmatic. By combining time courses of 4sU-seq, RNA-seq, ribosome profiling (RP), and RNA polymerase II (RNA Pol II) ChIP-seq during T cell activation, we illustrate genome-wide temporal dynamics for ∼10,000 genes. This approach reveals not only immediate-early and posttranscriptionally regulated genes but also coupled changes in transcription and translation for >90% of genes. Recruitment, rather than release of paused RNA Pol II, primarily mediates transcriptional changes. This coincides with a genome-wide temporary slowdown in cotranscriptional splicing, even for polyadenylated mRNAs that are localized at the chromatin. Subsequent splicing optimization correlates with increasing Ser-2 phosphorylation of the RNA Pol II carboxy-terminal domain (CTD) and activation of the positive transcription elongation factor (pTEFb). Thus, rapid de novo recruitment of RNA Pol II dictates the course of events during T cell activation, particularly transcription, splicing, and consequently translation. Display omitted •Genome-wide real-time expression analysis during a primary T helper response•Changes in transcription and translation are highly coupled during this response•On-time recruitment of RNA polymerase II dictates transcriptional changes•Cotranscriptional splicing rates temporarily drop at the beginning of the response Davari et al. visualize global changes in RNA Pol II binding, transcription, splicing, and translation. T cells change their functional program by rapid de novo recruitment of RNA Pol II and coupled changes in transcription and translation. This coincides with fluctuations in RNA Pol II phosphorylation and a temporary reduction in cotranscriptional splicing.
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