Transcription activation is enhanced by multivalent interactions independent of phase separation

Jorge Trojanowski; Lukas Frank; Anne Rademacher; Norbert Mücke; Pranas Grigaitis; Karsten Rippe

doi:10.1016/j.molcel.2022.04.017

Transcription activation is enhanced by multivalent interactions independent of phase separation

Jorge Trojanowski
, Lukas Frank
, Anne Rademacher
, Norbert Mücke
, Pranas Grigaitis
, Karsten Rippe^*

^*Corresponding author for this work

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Research output: Contribution to Journal › Article › Academic › peer-review

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Abstract

Transcription factors (TFs) consist of a DNA-binding domain and an activation domain (AD) that are frequently considered to be independent and exchangeable modules. However, recent studies report that the physicochemical properties of the AD can control TF assembly at chromatin by driving phase separation into transcriptional condensates. Here, we dissected transcription activation by comparing different synthetic TFs at a reporter gene array with real-time single-cell fluorescence microscopy. In these experiments, binding site occupancy, residence time, and coactivator recruitment in relation to multivalent TF interactions were compared. While phase separation propensity and activation strength of the AD were linked, the actual formation of liquid-like TF droplets had a neutral or inhibitory effect on transcription activation. We conclude that multivalent AD-mediated interactions enhance the transcription activation capacity of a TF by increasing its residence time in the chromatin-bound state and facilitating the recruitment of coactivators independent of phase separation.

Original language	English
Pages (from-to)	1878-1893.e10
Number of pages	27
Journal	Molecular cell
Volume	82
Issue number	10
Early online date	9 May 2022
DOIs	https://doi.org/10.1016/j.molcel.2022.04.017
Publication status	Published - 19 May 2022

Bibliographical note

Funding Information:
We thank Robin Weinmann and Fabian Erdel for discussions and the Bioquant advanced biological screening facility and the DKFZ light microscopy core facility for help. This project was supported by grant RI1283/16-1 in the Priority Program 2191 “Molecular Mechanisms of Functional Phase Separation” of the Deutsche Forschungsgemeinschaft (DFG) and the START-HD Explorer project of the University of Heidelberg . Data storage at SDS@hd was funded by the Ministry of Science, Research , and the Arts Baden-Württemberg (MWK) and the DFG through grants INST 35/1314-1 FUGG and INST 35/1503-1 FUGG.

Publisher Copyright:
© 2022 Elsevier Inc.

Funding

We thank Robin Weinmann and Fabian Erdel for discussions and the Bioquant advanced biological screening facility and the DKFZ light microscopy core facility for help. This project was supported by grant RI1283/16-1 in the Priority Program 2191 “Molecular Mechanisms of Functional Phase Separation” of the Deutsche Forschungsgemeinschaft (DFG) and the START-HD Explorer project of the University of Heidelberg . Data storage at SDS@hd was funded by the Ministry of Science, Research , and the Arts Baden-Württemberg (MWK) and the DFG through grants INST 35/1314-1 FUGG and INST 35/1503-1 FUGG.

Keywords

acetylation
BRD4
dCas9
gene regulation
kinetic proofreading
LLPS
multivalent interactions
optogenetics
phase separation
residence time
transcription factor
transcription kinetics

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10.1016/j.molcel.2022.04.017

Transcription activation is enhanced by multivalent interactions independent of phase separationFinal published version, 11 MBLicence: Article 25fa Dutch Copyright Act

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abstract = "Transcription factors (TFs) consist of a DNA-binding domain and an activation domain (AD) that are frequently considered to be independent and exchangeable modules. However, recent studies report that the physicochemical properties of the AD can control TF assembly at chromatin by driving phase separation into transcriptional condensates. Here, we dissected transcription activation by comparing different synthetic TFs at a reporter gene array with real-time single-cell fluorescence microscopy. In these experiments, binding site occupancy, residence time, and coactivator recruitment in relation to multivalent TF interactions were compared. While phase separation propensity and activation strength of the AD were linked, the actual formation of liquid-like TF droplets had a neutral or inhibitory effect on transcription activation. We conclude that multivalent AD-mediated interactions enhance the transcription activation capacity of a TF by increasing its residence time in the chromatin-bound state and facilitating the recruitment of coactivators independent of phase separation.",

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note = "Funding Information: We thank Robin Weinmann and Fabian Erdel for discussions and the Bioquant advanced biological screening facility and the DKFZ light microscopy core facility for help. This project was supported by grant RI1283/16-1 in the Priority Program 2191 “Molecular Mechanisms of Functional Phase Separation” of the Deutsche Forschungsgemeinschaft (DFG) and the START-HD Explorer project of the University of Heidelberg . Data storage at SDS@hd was funded by the Ministry of Science, Research , and the Arts Baden-W{\"u}rttemberg (MWK) and the DFG through grants INST 35/1314-1 FUGG and INST 35/1503-1 FUGG. Publisher Copyright: {\textcopyright} 2022 Elsevier Inc.",

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N2 - Transcription factors (TFs) consist of a DNA-binding domain and an activation domain (AD) that are frequently considered to be independent and exchangeable modules. However, recent studies report that the physicochemical properties of the AD can control TF assembly at chromatin by driving phase separation into transcriptional condensates. Here, we dissected transcription activation by comparing different synthetic TFs at a reporter gene array with real-time single-cell fluorescence microscopy. In these experiments, binding site occupancy, residence time, and coactivator recruitment in relation to multivalent TF interactions were compared. While phase separation propensity and activation strength of the AD were linked, the actual formation of liquid-like TF droplets had a neutral or inhibitory effect on transcription activation. We conclude that multivalent AD-mediated interactions enhance the transcription activation capacity of a TF by increasing its residence time in the chromatin-bound state and facilitating the recruitment of coactivators independent of phase separation.

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Transcription activation is enhanced by multivalent interactions independent of phase separation

Abstract

Bibliographical note

Funding

Keywords

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