Understanding p300-transcription factor interactions using sequence variation and hybridization

Fruzsina Hóbor; Zsófia Hegedüs; Amaurys Avila Ibarra; Vencel L. Petrovicz; Gail J. Bartlett; Richard B. Sessions; Andrew J. Wilson; Thomas A. Edwards

doi:10.1039/d2cb00026a

Understanding p300-transcription factor interactions using sequence variation and hybridization

Fruzsina Hóbor, Zsófia Hegedüs^*, Amaurys Avila Ibarra, Vencel L. Petrovicz, Gail J. Bartlett, Richard B. Sessions, Andrew J. Wilson^*, Thomas A. Edwards^*

^*Corresponding author for this work

Chemistry

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Abstract

The hypoxic response is central to cell function and plays a significant role in the growth and survival of solid tumours. HIF-1 regulates the hypoxic response by activating over 100 genes responsible for adaptation to hypoxia, making it a potential target for anticancer drug discovery. Although there is significant structural and mechanistic understanding of the interaction between HIF-1α and p300 alongside negative regulators of HIF-1α such as CITED2, there remains a need to further understand the sequence determinants of binding. In this work we use a combination of protein expression, chemical synthesis, fluorescence anisotropy and isothermal titration calorimetry for HIF-1α sequence variants and a HIF-1α-CITED hybrid sequence which we term CITIF. We show the HIF-1α sequence is highly tolerant to sequence variation through reduced enthalpic and less unfavourable entropic contributions, These data imply backbone as opposed to side chain interactions and ligand folding control the binding interaction and that sequence variations are tolerated as a result of adopting a more disordered bound interaction or “fuzzy” complex.

Original language	English
Pages (from-to)	592-603
Number of pages	12
Journal	RSC Chemical Biology
Volume	3
Issue number	5
Early online date	11 Apr 2022
DOIs	https://doi.org/10.1039/d2cb00026a
Publication status	Published - 1 May 2022

Bibliographical note

Funding:
This work was supported by EPSRC (EP/N013573/1 and EP/K039202/1) and the BBSRC/EPSRC-funded Synthetic Biology Research Centre, BrisSynBio (BB/L01386X/1). This project has received funding from the European Union's Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement no. MSCA-IF-2016-749012. Z.H. received funding from the National Research, Development and Innovation Office – NKFIH PD 135324. A. J. W. wishes to acknowledge the support of a Royal Society Leverhulme Trust Senior Fellowship (SRF\R1\191087).

Copyright:
© 2022 The Author(s). Published by the Royal Society of Chemistry.

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Biochemistry, Genetics and Molecular Biology (miscellaneous)
Chemistry (miscellaneous)

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Cite this

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title = "Understanding p300-transcription factor interactions using sequence variation and hybridization",

abstract = "The hypoxic response is central to cell function and plays a significant role in the growth and survival of solid tumours. HIF-1 regulates the hypoxic response by activating over 100 genes responsible for adaptation to hypoxia, making it a potential target for anticancer drug discovery. Although there is significant structural and mechanistic understanding of the interaction between HIF-1α and p300 alongside negative regulators of HIF-1α such as CITED2, there remains a need to further understand the sequence determinants of binding. In this work we use a combination of protein expression, chemical synthesis, fluorescence anisotropy and isothermal titration calorimetry for HIF-1α sequence variants and a HIF-1α-CITED hybrid sequence which we term CITIF. We show the HIF-1α sequence is highly tolerant to sequence variation through reduced enthalpic and less unfavourable entropic contributions, These data imply backbone as opposed to side chain interactions and ligand folding control the binding interaction and that sequence variations are tolerated as a result of adopting a more disordered bound interaction or “fuzzy” complex.",

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note = "Funding: This work was supported by EPSRC (EP/N013573/1 and EP/K039202/1) and the BBSRC/EPSRC-funded Synthetic Biology Research Centre, BrisSynBio (BB/L01386X/1). This project has received funding from the European Union's Horizon 2020 research and innovation programme under the Marie Sk{\l}odowska-Curie grant agreement no. MSCA-IF-2016-749012. Z.H. received funding from the National Research, Development and Innovation Office – NKFIH PD 135324. A. J. W. wishes to acknowledge the support of a Royal Society Leverhulme Trust Senior Fellowship (SRF\R1\191087). Copyright: {\textcopyright} 2022 The Author(s). Published by the Royal Society of Chemistry.",

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AU - Sessions, Richard B.

AU - Wilson, Andrew J.

AU - Edwards, Thomas A.

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Understanding p300-transcription factor interactions using sequence variation and hybridization

Abstract

Bibliographical note

ASJC Scopus subject areas

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