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Direct visualization of transcriptional activation by physical enhancer–promoter proximity

Hongtao Chen, Miki Fujioka, James B. Jaynes, Thomas Gregor
doi: https://doi.org/10.1101/099523
Hongtao Chen
aJoseph Henry Laboratories of Physics, and the Lewis–Sigler Institute for Integrative Genomics, Princeton University, Princeton NJ 08544, USA
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Miki Fujioka
bDepartment of Biochemistry and Molecular Biology, and the Kimmel Cancer Center, Thomas Jefferson University, Philadelphia PA 19107, USA
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James B. Jaynes
bDepartment of Biochemistry and Molecular Biology, and the Kimmel Cancer Center, Thomas Jefferson University, Philadelphia PA 19107, USA
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Thomas Gregor
aJoseph Henry Laboratories of Physics, and the Lewis–Sigler Institute for Integrative Genomics, Princeton University, Princeton NJ 08544, USA
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  • For correspondence: tg2@princeton.edu
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Abstract

A long-standing question in metazoan gene regulation is how remote enhancers communicate with their target promoters over long distances. Combining genome editing and quantitative live imaging we simultaneously visualize physical enhancer–promoter communication and transcription in Drosophila embryos. Enhancers regulating pair rule stripes of even-skipped expression activate transcription of a reporter gene over a distance of 150 kb. We show in individual cells that activation only occurs after the enhancer comes into close proximity with its regulatory target and that upon dissociation transcription ceases almost immediately. We further observe distinct topological conformations of the eve locus, depending on the spatial identity of the activating stripe enhancer. In addition, long-range activation results in transcriptional competition at the endogenous eve locus, causing corresponding developmental defects. Overall, we demonstrate that sustained physical proximity and enhancer–promoter engagement are required for enhancer action, and we provide a path to probe the implications of long-range regulation on cellular fates.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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Posted January 31, 2017.
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Direct visualization of transcriptional activation by physical enhancer–promoter proximity
Hongtao Chen, Miki Fujioka, James B. Jaynes, Thomas Gregor
bioRxiv 099523; doi: https://doi.org/10.1101/099523
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Direct visualization of transcriptional activation by physical enhancer–promoter proximity
Hongtao Chen, Miki Fujioka, James B. Jaynes, Thomas Gregor
bioRxiv 099523; doi: https://doi.org/10.1101/099523

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