RNA Polymerase II cluster dynamics predict mRNA output in living cells

Protein clustering is a hallmark of genome regulation in mammalian cells. However, the dynamic molecular processes involved make it difficult to correlate clustering with functional consequences in vivo. We developed a live-cell super-resolution approach to uncover the correlation between mRNA synth...

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Bibliographic Details
Published in:	eLife Vol. 5
Main Authors:	Cho, Won-Ki, Jayanth, Namrata, English, Brian P, Inoue, Takuma, Andrews, J Owen, Conway, William, Grimm, Jonathan B, Spille, Jan-Hendrik, Lavis, Luke D, Lionnet, Timothée, Cisse, Ibrahim I
Format:	Journal Article
Language:	English
Published:	England eLife Science Publications, Ltd 03-05-2016 eLife Sciences Publications Ltd eLife Sciences Publications, Ltd
Subjects:	Actin Actins - genetics Animals Biophysics and Structural Biology bursting Cameras Cell research Cells, Cultured Clustering DNA-directed RNA polymerase Embryo fibroblasts Embryos Fibroblasts - physiology Gene expression Gene Expression Regulation Genes and Chromosomes Genetic Loci Genetic regulation Genomes Hybridization Localization Mammalian cells Messenger RNA Mice Microscopy mRNA Proteins RNA polymerase RNA Polymerase II RNA Polymerase II - metabolism RNA polymerases RNA, Messenger - biosynthesis Transcription mouse transcription genes chromosomes clustering mRNA structural biology RNA Polymerase II biophysics bursting gene expression
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Summary:	Protein clustering is a hallmark of genome regulation in mammalian cells. However, the dynamic molecular processes involved make it difficult to correlate clustering with functional consequences in vivo. We developed a live-cell super-resolution approach to uncover the correlation between mRNA synthesis and the dynamics of RNA Polymerase II (Pol II) clusters at a gene locus. For endogenous β-actin genes in mouse embryonic fibroblasts, we observe that short-lived (~8 s) Pol II clusters correlate with basal mRNA output. During serum stimulation, a stereotyped increase in Pol II cluster lifetime correlates with a proportionate increase in the number of mRNAs synthesized. Our findings suggest that transient clustering of Pol II may constitute a pre-transcriptional regulatory event that predictably modulates nascent mRNA output.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	2050-084X 2050-084X
DOI:	10.7554/elife.13617