Allosteric activation mechanism of DriD, a WYL-domain containing transcription regulator

Abstract WYL-domain containing transcription factors regulate fundamental processes in bacterial physiology, yet how these proteins sense cellular cues to elicit an allosteric response is not well understood. Here we describe the allosteric activation mechanism of DriD, a Caulobacter crescentus homo...

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Main Authors: Emily Cannistraci, Bindu Y. Srinivasu, Jose Chavez Orozco, Kevin Gozzi, Thomas E. Wales, Maria A. Schumacher
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Communications Biology
Online Access:https://doi.org/10.1038/s42003-025-08111-x
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author Emily Cannistraci
Bindu Y. Srinivasu
Jose Chavez Orozco
Kevin Gozzi
Thomas E. Wales
Maria A. Schumacher
author_facet Emily Cannistraci
Bindu Y. Srinivasu
Jose Chavez Orozco
Kevin Gozzi
Thomas E. Wales
Maria A. Schumacher
author_sort Emily Cannistraci
collection DOAJ
description Abstract WYL-domain containing transcription factors regulate fundamental processes in bacterial physiology, yet how these proteins sense cellular cues to elicit an allosteric response is not well understood. Here we describe the allosteric activation mechanism of DriD, a Caulobacter crescentus homodimeric WYL-domain containing transcription regulator that activates a non-canonical DNA damage pathway. DriD senses ssDNA, produced upon DNA damage via interaction with its WYL domain. This stimulates DriD target DNA binding. However, its DNA-binding domains (DNABDs) are 50 Å from the WYL-domains and linked by a three-helix bundle domain (3HB). Using a combination of crystallography, biochemistry, and HDX-MS we unveil an allosteric mechanism whereby an inhibitory interaction, formed between the DriD DNABD and 3HB in the apo form, is freed upon ssDNA binding, allowing target DNA binding. These findings may serve as a model for understanding activation by the large family of homodimeric WYL activators, including those in pathogenic bacteria.
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spelling doaj-art-e0a747167de8443a8f9563dfa40617a22025-08-20T01:47:29ZengNature PortfolioCommunications Biology2399-36422025-04-018111510.1038/s42003-025-08111-xAllosteric activation mechanism of DriD, a WYL-domain containing transcription regulatorEmily Cannistraci0Bindu Y. Srinivasu1Jose Chavez Orozco2Kevin Gozzi3Thomas E. Wales4Maria A. Schumacher5Department of Biochemistry, 307 Research Dr., Box 3711, Duke University Medical CenterDepartment of Chemistry and Chemical Biology, Northeastern UniversityDepartment of Biochemistry, 307 Research Dr., Box 3711, Duke University Medical Center100 Edwin H Land Blvd, Harvard University CambridgeDepartment of Chemistry and Chemical Biology, Northeastern UniversityDepartment of Biochemistry, 307 Research Dr., Box 3711, Duke University Medical CenterAbstract WYL-domain containing transcription factors regulate fundamental processes in bacterial physiology, yet how these proteins sense cellular cues to elicit an allosteric response is not well understood. Here we describe the allosteric activation mechanism of DriD, a Caulobacter crescentus homodimeric WYL-domain containing transcription regulator that activates a non-canonical DNA damage pathway. DriD senses ssDNA, produced upon DNA damage via interaction with its WYL domain. This stimulates DriD target DNA binding. However, its DNA-binding domains (DNABDs) are 50 Å from the WYL-domains and linked by a three-helix bundle domain (3HB). Using a combination of crystallography, biochemistry, and HDX-MS we unveil an allosteric mechanism whereby an inhibitory interaction, formed between the DriD DNABD and 3HB in the apo form, is freed upon ssDNA binding, allowing target DNA binding. These findings may serve as a model for understanding activation by the large family of homodimeric WYL activators, including those in pathogenic bacteria.https://doi.org/10.1038/s42003-025-08111-x
spellingShingle Emily Cannistraci
Bindu Y. Srinivasu
Jose Chavez Orozco
Kevin Gozzi
Thomas E. Wales
Maria A. Schumacher
Allosteric activation mechanism of DriD, a WYL-domain containing transcription regulator
Communications Biology
title Allosteric activation mechanism of DriD, a WYL-domain containing transcription regulator
title_full Allosteric activation mechanism of DriD, a WYL-domain containing transcription regulator
title_fullStr Allosteric activation mechanism of DriD, a WYL-domain containing transcription regulator
title_full_unstemmed Allosteric activation mechanism of DriD, a WYL-domain containing transcription regulator
title_short Allosteric activation mechanism of DriD, a WYL-domain containing transcription regulator
title_sort allosteric activation mechanism of drid a wyl domain containing transcription regulator
url https://doi.org/10.1038/s42003-025-08111-x
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