Twist-dependent ratchet functioning downstream from Dorsal revealed using a light-inducible degron
Abstract
Graded transcription factors are pivotal regulators of embryonic patterning, but whether their role changes over time is unclear. A light-regulated protein degradation system was used to assay temporal dependence of the transcription factor Dorsal in dorsal–ventral axis patterning of Drosophila embryos. Surprisingly, the high-threshold target gene snail only requires Dorsal input early but not late when Dorsal levels peak. Instead, late snail expression can be supported by action of the Twist transcription factor, specifically, through one enhancer, sna.distal. This study demonstrates that continuous input is not required for some Dorsal targets and downstream responses, such as twist, function as molecular ratchets.
Additional Information
© 2020 Irizarry et al.; Published by Cold Spring Harbor Laboratory Press This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. Received March 12, 2020; revised version accepted April 24, 2020. Published in Advance May 28, 2020. We are grateful to Mike Levine and Hernan Garcia for providing fly stocks, Leslie Dunipace for injections and guidance, Theodora Koromila for help with imaging strategy, and Susan Newcomb for edits to manuscript. This study was supported by funding from National Institutes of Health grants R21HD095639 and R35GM118146 to A.S. and R21OD017964 to D.S. Author contributions: A.S., D.S., J.I., and J.M. conceived the project and planned the experimental approach. A.S. directed the project. J.I. performed all imaging. J.M. performed all quantitative analysis of the imaging data, Crispr/Cas9 genomic engineering, Western analysis, and viability studies. J.I. and J.M. performed stainings. D.S. and G.K. validated the use of BLID for studies in Drosophila and helped to develop protocols for its use. J.I., J.M., and D.S. developed the protocol for fixed embryo analysis, whereas J.I. and J.M. developed protocols for BLID live imaging.Data were analyzed by J.I., J.M., and A.S. The manuscript was written by J.I., J.M., and A.S. with edits provided by D.S.Attached Files
Published - Genes_Dev.-2020-Irizarry-965-72.pdf
Supplemental Material - Supplemental_Material.docx
Supplemental Material - Supplemental_Movie_S1.mov
Supplemental Material - Supplemental_Movie_S10.mov
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Supplemental Material - Supplemental_Movie_S6.mov
Supplemental Material - Supplemental_Movie_S7.mov
Supplemental Material - Supplemental_Movie_S8.mov
Supplemental Material - Supplemental_Movie_S9.mov
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Additional details
- PMCID
- PMC7328519
- Eprint ID
- 103542
- DOI
- 10.1101/gad.338194.120
- Resolver ID
- CaltechAUTHORS:20200529-090635112
- R21HD095639
- NIH
- R35GM118146
- NIH
- R21OD017964
- NIH
- Created
-
2020-05-29Created from EPrint's datestamp field
- Updated
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2022-07-05Created from EPrint's last_modified field
- Caltech groups
- Division of Biology and Biological Engineering