Autoinhibition and Activation Mechanisms for MICAL Monooxygenases in F-actin Disassembly

Autoinhibition and Activation Mechanisms for MICAL Monooxygenases in F-actin Disassembly | 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 Autoinhibition and Activation Mechanisms for MICAL Monooxygenases in F-actin Disassembly Zhiyi Wei, Leishu Lin, Jiayuan Dong, Shun Xu, Jinman Xiao, Fengfeng Niu, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3960862/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 09 Aug, 2024 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract MICAL (Molecule Interacting with CasL) proteins represent a unique family of actin regulators crucial for synapse development, membrane trafficking, and cytokinesis. Unlike classical actin regulators, MICAL proteins possess enzymatic activity, catalyzing the oxidation of specific residues within actin filaments to induce robust filament severing. The potent activity of MICAL proteins requires tight control to prevent extensive damage to the actin cytoskeleton. Due to limited structural information on full-length MICAL proteins, the molecular mechanisms governing MICAL autoinhibition and activation remain elusive. Here, we reported the cryo-EM structure of full-length MICAL1, unveiling a head-to-tail interaction that allosterically blocks enzymatic activity through the binding of the C-terminal Rab-binding domain (RBD) to the N-terminal monooxygenase domain. The structure also reveals the assembly of C-terminal domains via a tripartite interdomain interaction, stabilizing the inhibitory conformation of the RBD. Our structural, biochemical, and cellular analyses elucidate the transition from the autoinhibited to activated conformation of MICAL1 in response to Rab8-subfamily GTPase. Dual Rab-binding to the RBD induces the conformation rearrangement of the RBD and synergistically promotes the autoinhibition-to-activation transition, revealing intricate activity regulation mechanisms for MICAL proteins. Furthermore, our mutagenesis study of MICAL3 suggests a conserved autoinhibition and activation mechanism among MICAL proteins. Biological sciences/Biochemistry/Structural biology/Electron microscopy/Cryoelectron microscopy Biological sciences/Cell biology/Cytoskeleton/Actin MICAL-1 MICAL family cytoskeleton F-actin depolymerization Rab Full Text Additional Declarations There is NO Competing Interest. Supplementary Files MovieS1.mp4 Supplementary Movie 1 Cite Share Download PDF Status: Published Journal Publication published 09 Aug, 2024 Read the published version in Nature Communications → 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-3960862","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":274501252,"identity":"7b414eb2-9adc-42a2-b209-62cca8885125","order_by":0,"name":"Zhiyi 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Unlike classical actin regulators, MICAL proteins possess enzymatic activity, catalyzing the oxidation of specific residues within actin filaments to induce robust filament severing. The potent activity of MICAL proteins requires tight control to prevent extensive damage to the actin cytoskeleton. Due to limited structural information on full-length MICAL proteins, the molecular mechanisms governing MICAL autoinhibition and activation remain elusive. Here, we reported the cryo-EM structure of full-length MICAL1, unveiling a head-to-tail interaction that allosterically blocks enzymatic activity through the binding of the C-terminal Rab-binding domain (RBD) to the N-terminal monooxygenase domain. The structure also reveals the assembly of C-terminal domains via a tripartite interdomain interaction, stabilizing the inhibitory conformation of the RBD. Our structural, biochemical, and cellular analyses elucidate the transition from the autoinhibited to activated conformation of MICAL1 in response to Rab8-subfamily GTPase. Dual Rab-binding to the RBD induces the conformation rearrangement of the RBD and synergistically promotes the autoinhibition-to-activation transition, revealing intricate activity regulation mechanisms for MICAL proteins. 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