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Dynamic multifactor hubs interact transiently with sites of active transcription in Drosophila embryos

View ORCID ProfileMustafa Mir, View ORCID ProfileMichael R. Stadler, Stephan A. Ortiz, View ORCID ProfileMelissa M. Harrison, View ORCID ProfileXavier Darzacq, View ORCID ProfileMichael B. Eisen
doi: https://doi.org/10.1101/377812
Mustafa Mir
1Department of Molecular and Cell Biology, University of California, Berkeley, CA, 94720 U.S.A.
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Michael R. Stadler
1Department of Molecular and Cell Biology, University of California, Berkeley, CA, 94720 U.S.A.
2Howard Hughes Medical Institute, University of California, Berkeley, CA, 94720 U.S.A.
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Stephan A. Ortiz
1Department of Molecular and Cell Biology, University of California, Berkeley, CA, 94720 U.S.A.
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Melissa M. Harrison
3Department of Biomolecular Chemistry, University of Wisconsin, Madison, WI 53706 U.S.A.
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Xavier Darzacq
1Department of Molecular and Cell Biology, University of California, Berkeley, CA, 94720 U.S.A.
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  • For correspondence: mbeisen@berkeley.edu darzacq@berkeley.edu
Michael B. Eisen
1Department of Molecular and Cell Biology, University of California, Berkeley, CA, 94720 U.S.A.
2Howard Hughes Medical Institute, University of California, Berkeley, CA, 94720 U.S.A.
4Department of Integrative Biology, University of California, Berkeley, CA, 94720 U.S.A.
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  • For correspondence: mbeisen@berkeley.edu darzacq@berkeley.edu
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Abstract

The regulation of transcription requires the coordination of numerous activities on DNA, yet it remains poorly understood how transcription factors facilitate these multiple functions. Here we use lattice light-sheet microscopy to integrate single-molecule and high-speed 4D imaging in developing Drosophila embryos to study the nuclear organization and interactions of the key patterning factors Zelda and Bicoid. In contrast to previous studies suggesting stable, cooperative binding, we show that both factors interact with DNA with surprisingly high off-rates. We find that both factors form dynamic subnuclear hubs, and that Bicoid binding is enriched within Zelda hubs. Remarkably, these hubs are both short lived and interact only transiently with sites of active Bicoid dependent transcription. Based on our observations we hypothesize that, beyond simply forming bridges between DNA and the transcription machinery, transcription factors can organize other proteins into hubs that transiently drive multiple activities at their gene targets.

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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 4.0 International license.
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Posted July 26, 2018.
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Dynamic multifactor hubs interact transiently with sites of active transcription in Drosophila embryos
Mustafa Mir, Michael R. Stadler, Stephan A. Ortiz, Melissa M. Harrison, Xavier Darzacq, Michael B. Eisen
bioRxiv 377812; doi: https://doi.org/10.1101/377812
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Dynamic multifactor hubs interact transiently with sites of active transcription in Drosophila embryos
Mustafa Mir, Michael R. Stadler, Stephan A. Ortiz, Melissa M. Harrison, Xavier Darzacq, Michael B. Eisen
bioRxiv 377812; doi: https://doi.org/10.1101/377812

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