Transcription activation mechanism of a non-canonical bacterial DNA damage response pathway

Transcription activation mechanism of a non-canonical bacterial DNA damage response pathway | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Transcription activation mechanism of a non-canonical bacterial DNA damage response pathway Maria Schumacher, Rajiv Singh, Amani Chinni, Emily Cannistraci, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7152246/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract DNA damage repair mechanisms are vital for bacterial survival. Recent studies revealed a non-canonical DNA damage response in Caulobacter crescentus activated by a WYL-domain transcription factor, DriD. DriD binds ssDNA, produced upon DNA damage, within its WYL-domain, and drives expression at multiple promoters. The mechanism behind DriD-mediated transcription activation is, however, unknown. Here we describe cryo-EM structures of DriD-ssDNA bound to RNAP-holoenzyme and three promoters. DriD contains N-terminal DNA-binding domains (DNABDs) connected to WYL-signaling domains by a linker-3-helix-bundle (3HB) module. The three structures reveal a conserved activation mechanism whereby DriD’s 3HBs bind RNAP α-CTD and ß domains, anchoring RNAP on nonoptimal promoters. The 3HBs form autoinhibitory contacts with DNABDs in apo DriD and therefore acts as a ssDNA-driven trigger domain, switching between DNABD-bound apo and RNAP-bound states upon ssDNA-mediated activation. Thus, the structures reveal a unique transcription activation mechanism, likely conserved among the large family of homodimeric WYL-activators. Biological sciences/Structural biology/Electron microscopy/Cryoelectron microscopy Biological sciences/Microbiology/Bacteria/Bacterial structural biology Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SinghSupplementatyMatMicroJuly12.pdf Supplementary Information SupplementaryMovieFiguresLegends.docx Supplementary Movie 1: Figure legend SupplementaryMovie1.mp4 Supplementary Movie 1 D1000297728valreportfullannotateP1.pdf PDB validation report pdbfiles.zip structure models Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7152246","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":495558603,"identity":"08116621-6a74-4f43-994d-276a331024e7","order_by":0,"name":"Maria 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