Molecular switching in transcription through splicing and proline-isomerization regulates stress responses in plants

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The Arabidopsis thaliana DREB2A transcription factor interacts with the negative regulator RCD1 and the ACID domain of subunit 25 of the transcriptional co-regulator mediator (Med25) to integrate stress signals for gene expression, with elusive molecular interplay. Using biophysical and structural analyses together with high-throughput screening, we reveal a bivalent binding switch in DREB2A containing an ACID-binding motif (ABS) and the known RCD1-binding motif (RIM). The RIM is lacking in a stress-induced DREB2A splice variant with retained transcriptional activity. ABS and RIM bind to separate sites on Med25-ACID, and NMR analyses show a structurally heterogeneous complex deriving from a DREB2A-ABS proline residue populating cis- and trans-isomers with remote impact on the RIM. The cis-isomer stabilizes an α-helix, while the trans-isomer may introduce energetic frustration facilitating rapid exchange between activators and repressors. Thus, DREB2A uses a post-transcriptionally and post-translationally modulated switch for transcriptional regulation.
OriginalsprogEngelsk
Artikelnummer592
TidsskriftNature Communications
Vol/bind15
Udgave nummer1
Antal sider13
ISSN2041-1723
DOI
StatusUdgivet - 2024

Bibliografisk note

Funding Information:
This work was supported by the Novo Nordisk Foundation challenge grant REPIN, rethinking protein interactions (grant no.: NNF18OC0033926; to B.B.K. and K.S.) and by the Danish Research Council (grant no.: 9040-00164B to B.B.K. and K.S.). Activation domain assay was done under the National Science Foundation (NSF) (PGRP BIO-2112056 to L.C.S.; Postdoctoral Research Program IOS-1907098 to N.M.). NMR spectra were recorded at cOpenNMR, an infrastructure supported by the Novo Nordisk Foundation (grant no.: NNF18OC0032996). The authors would like to thank Dr. Anders Lønstrup Hansen for many important discussions regarding analysis and representation of data. We also thank Prof. Lewis E. Kay for sharing the C-CEST pulse sequences necessary for this work, Dr. Elise Delaforge for discussions and suggestions regarding analysis of CEST data and Stase Bielskute for suggesting the δ2D tool for analysis of IDP secondary structure. We wish to thank Cy Jeffries at the EMBL Hamburg beamline for recording, processing and providing initial analysis of SAXS data. We thank Profs. Ben Schuler and Per Jemth for valuable discussions of kinetics data leading to further exploration. 13

Publisher Copyright:
© 2024, The Author(s).

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