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Elife. 2018 Dec 27;7. pii: e40497. doi: 10.7554/eLife.40497.

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

Author information

1
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States.
2
Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States.
3
Department of Biomolecular Chemistry, University of Wisconsin-Madison, Madison, United States.
4
Department of Integrative Biology, University of California, Berkeley, Berkeley, United States.
#
Contributed equally

Abstract

The regulation of transcription requires the coordination of numerous activities on DNA, yet how transcription factors mediate these activities remains poorly understood. 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 transcription 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.

Editorial note:

This article has been through an editorial process in which the authors decide how to respond to the issues raised during peer review. The Reviewing Editor's assessment is that all the issues have been addressed (see decision letter).

KEYWORDS:

D. melanogaster; bicoid; chromosomes; developmental biology; gene expression; membraneless compartments; ms2; single molecule imaging; transcription regulation; zelda

PMID:
30589412
PMCID:
PMC6307861
DOI:
10.7554/eLife.40497
[Indexed for MEDLINE]
Free PMC Article

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