RNA polymerase II clustering through CTD phase separation

Marc Boehning; Claire Dugast‐Darzacq; Marija Rankovic; Anders S. Hansen; Taekyung Yu; Hervé Marie-Nelly; David Trombley McSwiggen; Goran Kokić; Gina M. Dailey; Patrick Cramer; Xavier Darzacq; Markus Zweckstetter

doi:10.1038/s41594-018-0112-y

Molecular BiologyNature Structural & Molecular BiologyAug 10, 2018

Peer-reviewed journal article

RNA polymerase II clustering through CTD phase separation

Marc Boehning, Claire Dugast‐Darzacq iD, Marija Rankovic, Anders S. Hansen iD, Taekyung Yu iD, Hervé Marie-Nelly iD, David Trombley McSwiggen iD, Goran Kokić iD, Gina M. Dailey iD, Patrick Cramer iD, Xavier Darzacq iD, Markus Zweckstetter iD

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Abstract

The carboxy-terminal domain (CTD) of RNA polymerase (Pol) II is an intrinsically disordered low-complexity region that is critical for pre-mRNA transcription and processing. The CTD consists of hepta-amino acid repeats varying in number from 52 in humans to 26 in yeast. Here we report that human and yeast CTDs undergo cooperative liquid phase separation at increasing protein concentration, with the shorter yeast CTD forming less stable droplets. In human cells, truncation of the CTD to the length of the yeast CTD decreases Pol II clustering and chromatin association whereas CTD extension has the opposite effect. CTD droplets can incorporate intact Pol II and are dissolved by CTD phosphorylation with the transcription initiation factor IIH kinase CDK7. Together with published data, our results suggest that Pol II forms clusters/hubs at active genes through interactions between CTDs and with activators, and that CTD phosphorylation liberates Pol II enzymes from hubs for promoter escape and transcription elongation.