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Single-molecule imaging reveals the interplay between transcription factors, nucleosomes, and transcriptional bursting

Benjamin T. Donovan, Anh Huynh, David A. Ball, Michael G. Poirier, Daniel R. Larson, Matthew L. Ferguson, Tineke L. Lenstra
doi: https://doi.org/10.1101/404681
Benjamin T. Donovan
1Biophysics Graduate Program, The Ohio State University, Columbus, Ohio 43210, USA
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Anh Huynh
2Department of Physics, Boise State University, Boise, Idaho 83725, USA.
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David A. Ball
3Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, NIH, Bethesda, MD 20892, USA
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Michael G. Poirier
1Biophysics Graduate Program, The Ohio State University, Columbus, Ohio 43210, USA
4Department of Physics, Ohio State Biochemistry Program, The Ohio State University, Columbus, Ohio 43210, USA
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Daniel R. Larson
3Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, NIH, Bethesda, MD 20892, USA
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Matthew L. Ferguson
2Department of Physics, Boise State University, Boise, Idaho 83725, USA.
5Biomolecular Sciences, Boise State University, Boise, Idaho 83725, USA.
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  • For correspondence: mattferguson@boisestate.edu t.lenstra@nki.nl
Tineke L. Lenstra
6Division of Gene Regulation, the Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands
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  • For correspondence: mattferguson@boisestate.edu t.lenstra@nki.nl
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Summary

Transcription factors show rapid and reversible binding to chromatin in living cells, and transcription occurs in sporadic bursts, but how these phenomena are related is unknown. Using a combination of in vitro and in vivo single-molecule imaging approaches, we directly correlated binding of the transcription factor Gal4 with the transcriptional bursting kinetics of the Gal4 target genes GAL3 and GAL10 in living yeast cells. We find that Gal4 dwell times sets the transcriptional burst size. Gal4 dwell time depends on the affinity of the binding site and is reduced by orders of magnitude by nucleosomes. Using a novel imaging platform, we simultaneously tracked transcription factor binding and transcription at one locus, revealing the timing and correlation between Gal4 binding and transcription. Collectively, our data support a model where multiple polymerases initiate during a burst as long as the transcription factor is bound to DNA, and a burst terminates upon transcription factor dissociation.

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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-ND 4.0 International license.
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Posted August 31, 2018.
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Single-molecule imaging reveals the interplay between transcription factors, nucleosomes, and transcriptional bursting
Benjamin T. Donovan, Anh Huynh, David A. Ball, Michael G. Poirier, Daniel R. Larson, Matthew L. Ferguson, Tineke L. Lenstra
bioRxiv 404681; doi: https://doi.org/10.1101/404681
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Single-molecule imaging reveals the interplay between transcription factors, nucleosomes, and transcriptional bursting
Benjamin T. Donovan, Anh Huynh, David A. Ball, Michael G. Poirier, Daniel R. Larson, Matthew L. Ferguson, Tineke L. Lenstra
bioRxiv 404681; doi: https://doi.org/10.1101/404681

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