The WG and DPP morphogens regulate gene expression by modulating the frequency of transcriptional bursts

Rachael Bakker; Madhav Mani; Richard W. Carthew

doi:10.7554/eLife.56076

The WG and DPP morphogens regulate gene expression by modulating the frequency of transcriptional bursts

Rachael Bakker, Madhav Mani^*, Richard W. Carthew^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

Morphogen signaling contributes to the patterned spatiotemporal expression of genes during development. One mode of regulation of signaling-responsive genes is at the level of transcription. Single-cell quantitative studies of transcription have revealed that transcription occurs intermittently, in bursts. Although the effects of many gene regulatory mechanisms on transcriptional bursting have been studied, it remains unclear how morphogen gradients affect this dynamic property of downstream genes. Here we have adapted single molecule fluorescence in situ hybridization (smFISH) for use in the Drosophila wing imaginal disc in order to measure nascent and mature mRNA of genes downstream of the Wg and Dpp morphogen gradients. We compared our experimental results with predictions from stochastic models of transcription, which indicated that the transcription levels of these genes appear to share a common method of control via burst frequency modulation. Our data help further elucidate the link between developmental gene regulatory mechanisms and transcriptional bursting.

Original language	English (US)
Article number	e56076
Pages (from-to)	1-26
Number of pages	26
Journal	eLife
Volume	9
DOIs	https://doi.org/10.7554/eLife.56076
State	Published - Jun 2020

Funding

Fly stocks from Hugo Bellen and the Bloomington Drosophila Stock Center are gratefully appreciated. We thank Jessica Hornick and the Biological Imaging Facility for help with imaging and the Keck Facility at Northwestern for help with probe purification. We are very grateful to Shawn Little and Thomas Gregor for hosting RB at Princeton and their invaluable advice on adapting the smFISH method to imaginal discs. We also thank Arjun Raj and Brian Munsky for key suggestions on experimental and analytical development. Financial support was provided from the NIH (T32CA080621, RB; R35GM118144, RWC), NSF (1764421, MM and RWC), and the Simons Foundation (597491, MM and RWC). MM is a Simons Foundation Investigator.

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

Access to Document

10.7554/eLife.56076

Cite this

@article{b7d8f2f110a04d14905e97452bc32949,

title = "The WG and DPP morphogens regulate gene expression by modulating the frequency of transcriptional bursts",

abstract = "Morphogen signaling contributes to the patterned spatiotemporal expression of genes during development. One mode of regulation of signaling-responsive genes is at the level of transcription. Single-cell quantitative studies of transcription have revealed that transcription occurs intermittently, in bursts. Although the effects of many gene regulatory mechanisms on transcriptional bursting have been studied, it remains unclear how morphogen gradients affect this dynamic property of downstream genes. Here we have adapted single molecule fluorescence in situ hybridization (smFISH) for use in the Drosophila wing imaginal disc in order to measure nascent and mature mRNA of genes downstream of the Wg and Dpp morphogen gradients. We compared our experimental results with predictions from stochastic models of transcription, which indicated that the transcription levels of these genes appear to share a common method of control via burst frequency modulation. Our data help further elucidate the link between developmental gene regulatory mechanisms and transcriptional bursting.",

author = "Rachael Bakker and Madhav Mani and Carthew, \{Richard W.\}",

note = "Publisher Copyright: {\textcopyright} Bakker et al.",

year = "2020",

month = jun,

doi = "10.7554/eLife.56076",

language = "English (US)",

volume = "9",

pages = "1--26",

journal = "eLife",

issn = "2050-084X",

publisher = "eLife Sciences Publications Ltd",

}

TY - JOUR

T1 - The WG and DPP morphogens regulate gene expression by modulating the frequency of transcriptional bursts

AU - Bakker, Rachael

AU - Mani, Madhav

AU - Carthew, Richard W.

N1 - Publisher Copyright: © Bakker et al.

PY - 2020/6

Y1 - 2020/6

N2 - Morphogen signaling contributes to the patterned spatiotemporal expression of genes during development. One mode of regulation of signaling-responsive genes is at the level of transcription. Single-cell quantitative studies of transcription have revealed that transcription occurs intermittently, in bursts. Although the effects of many gene regulatory mechanisms on transcriptional bursting have been studied, it remains unclear how morphogen gradients affect this dynamic property of downstream genes. Here we have adapted single molecule fluorescence in situ hybridization (smFISH) for use in the Drosophila wing imaginal disc in order to measure nascent and mature mRNA of genes downstream of the Wg and Dpp morphogen gradients. We compared our experimental results with predictions from stochastic models of transcription, which indicated that the transcription levels of these genes appear to share a common method of control via burst frequency modulation. Our data help further elucidate the link between developmental gene regulatory mechanisms and transcriptional bursting.

AB - Morphogen signaling contributes to the patterned spatiotemporal expression of genes during development. One mode of regulation of signaling-responsive genes is at the level of transcription. Single-cell quantitative studies of transcription have revealed that transcription occurs intermittently, in bursts. Although the effects of many gene regulatory mechanisms on transcriptional bursting have been studied, it remains unclear how morphogen gradients affect this dynamic property of downstream genes. Here we have adapted single molecule fluorescence in situ hybridization (smFISH) for use in the Drosophila wing imaginal disc in order to measure nascent and mature mRNA of genes downstream of the Wg and Dpp morphogen gradients. We compared our experimental results with predictions from stochastic models of transcription, which indicated that the transcription levels of these genes appear to share a common method of control via burst frequency modulation. Our data help further elucidate the link between developmental gene regulatory mechanisms and transcriptional bursting.

UR - http://www.scopus.com/inward/record.url?scp=85087959726&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85087959726&partnerID=8YFLogxK

U2 - 10.7554/eLife.56076

DO - 10.7554/eLife.56076

M3 - Article

C2 - 32568073

AN - SCOPUS:85087959726

SN - 2050-084X

VL - 9

SP - 1

EP - 26

JO - eLife

JF - eLife

M1 - e56076

ER -

The WG and DPP morphogens regulate gene expression by modulating the frequency of transcriptional bursts

Abstract

Funding

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this