Family-wide analysis of human macrodomains reveals novel activities

Family-wide analysis of human macrodomains reveals novel activities | 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 Family-wide analysis of human macrodomains reveals novel activities Lisa Weixler*, Roko Žaja*, Nonso J. Ikenga, Ganga Mohan, Gülcan Aydin, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4644671/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 18 Mar, 2025 Read the published version in Communications Biology → Version 1 posted You are reading this latest preprint version Abstract ADP-ribosylation is well-known as protein posttranslational modification and was recently also identified as RNA posttranscriptional modification. ADP-ribose is added onto substrates by PARP enzymes and removed by the structurally distinct ADP-ribosylhydrolases (ARH) or macrodomain-containing proteins. When macrodomain proteins were identified as hydrolases a decade ago, many ADP-ribosylation substrates were not yet identified. Therefore, the majority of macrodomain-containing proteins have not been tested towards these additional substrates and were considered to be inactive. Here, we compare in vitro activities of the human macrodomain-containing proteins on a wide range of ADP-ribosylated substrates. We confirm recent findings that PARP9 macro1 and PARP14macro1 can reverse ADP-ribosylation from acidic residues and provide evidence that also PARP14macro2 and PARP15macro2 can function as ADP-ribosylhydrolases. In addition we identified both PARP9macro1 and PARP14macro1 as RNA decapping protein domains. Notwithstanding these in vitro activities, our data further show that PARG is the major RNA decapping enzyme in HEK293 cells. Together, our findings expand on defining catalytic functions of macrodomains including some previously thought to be only readers. *Lisa Weixler and Roko Žaja are shared first authors. Biological sciences/Biochemistry/Enzymes/Hydrolases Biological sciences/Molecular biology/Post-translational modifications/PolyADP-ribosylation Biological sciences/Cell biology/Post-translational modifications/PolyADP-ribosylation Full Text Additional Declarations There is NO Competing Interest. Cite Share Download PDF Status: Published Journal Publication published 18 Mar, 2025 Read the published version in Communications Biology → 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. 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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-4644671","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":324664932,"identity":"85b794b9-9fd0-424d-9de0-99b19b3fa00a","order_by":0,"name":"Lisa Weixler*","email":"","orcid":"https://orcid.org/0000-0003-4453-017X","institution":"Institute of Biochemistry and Molecular Biology, Pauwelsstraße 30, RWTH Aachen University, Germany; Institute for Clinical Chemistry and Clinical Pharmacology, Venusberg-Campus 1, University Hospital Bonn, Germany","correspondingAuthor":false,"prefix":"","firstName":"Lisa","middleName":"","lastName":"Weixler*","suffix":""},{"id":324664933,"identity":"8735c5c5-1c75-4494-938d-7a98d30b9cfd","order_by":1,"name":"Roko Žaja*","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA0klEQVRIiWNgGAWjYJACA4YKIGZgYANxGBuI03KGVC0MjG2kaDHvP/yg4Oe8w8b87YefPfi4g0G2n5AWmQPHDAx7tx02kziTZm448wyD8UxC1kgwNhgY8G47bMNwIIdNmreNIXHDAUJamNk/GP6dc9hG/vwbNum/QC37CWph4zEw5m04bGZwA2gLI8gWQn6R4OEpMJY5lm5seOOZmWRvm4TxDIK28B/fZvimxtpw3vnkZxI/22xk+xsIWQOMDgNkIwirBwLmB0QpGwWjYBSMgpELACU5PoQgMCMYAAAAAElFTkSuQmCC","orcid":"","institution":"Institute of Biochemistry and Molecular Biology, Pauwelsstraße 30, RWTH Aachen University, Germany","correspondingAuthor":true,"prefix":"","firstName":"Roko","middleName":"","lastName":"Žaja*","suffix":""},{"id":324664934,"identity":"5967c6e7-cda6-4d3f-9f82-bebae291a407","order_by":2,"name":"Nonso J. 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