Mediator and RNA polymerase II clusters associate in transcription-dependent condensates

Models of gene control have emerged from genetic and biochemical studies, with limited consideration of the spatial organization and dynamics of key components in living cells. We used live-cell superresolution and light-sheet imaging to study the organization and dynamics of the Mediator coactivato...

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Published in:	Science (American Association for the Advancement of Science) Vol. 361; no. 6400; pp. 412 - 415
Main Authors:	Cho, Won-Ki, Spille, Jan-Hendrik, Hecht, Micca, Lee, Choongman, Li, Charles, Grube, Valentin, Cisse, Ibrahim I
Format:	Journal Article
Language:	English
Published:	United States The American Association for the Advancement of Science 27-07-2018
Subjects:	Amino acid sequence Animals Cells, Cultured Chromatin Chromatin - metabolism Clusters Cofactors Complexity Condensates DNA-directed RNA polymerase Elongation Embryo cells Embryonic Stem Cells - metabolism Enhancer Elements, Genetic Enhancers Gene expression Gene Expression Regulation Gene regulation Hubs Inhibitors Luminescent Proteins - analysis Luminescent Proteins - chemistry Mediator Complex - analysis Mediator Complex - chemistry Mediator Complex - metabolism Mice Molecular Imaging - methods Phase separation Protein interaction Proteins Ribonucleic acid RNA RNA polymerase RNA polymerase II RNA Polymerase II - analysis RNA Polymerase II - chemistry RNA Polymerase II - metabolism Stem cells Transcription elongation Transcription factors Transcription Factors - metabolism Transcription, Genetic
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Summary:	Models of gene control have emerged from genetic and biochemical studies, with limited consideration of the spatial organization and dynamics of key components in living cells. We used live-cell superresolution and light-sheet imaging to study the organization and dynamics of the Mediator coactivator and RNA polymerase II (Pol II) directly. Mediator and Pol II each form small transient and large stable clusters in living embryonic stem cells. Mediator and Pol II are colocalized in the stable clusters, which associate with chromatin, have properties of phase-separated condensates, and are sensitive to transcriptional inhibitors. We suggest that large clusters of Mediator, recruited by transcription factors at large or clustered enhancer elements, interact with large Pol II clusters in transcriptional condensates in vivo.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 equal contribution Author contributions: W.K.C., J.-H.S., and I.I.C. conceived and designed the study; W.K.C. and J.-H.S. performed experiments and analyzed data with help from M.H., C.Lee, and V.G.; M.H. cloned CRISPR repair templates and sgRNA plasmids and genotyped cell lines; C.Li conducted and analyzed the Western Blot and ChIP-seq assay. W.K.C., J.-H.S., and I.I.C. wrote the manuscript with input from all co-authors. I.I.C. supervised all aspects of the project.
ISSN:	0036-8075 1095-9203
DOI:	10.1126/science.aar4199