{"oa":1,"publisher":"Elsevier","file_date_updated":"2023-01-24T09:29:02Z","doi":"10.1016/j.molcel.2022.10.010","file":[{"file_size":7368534,"date_created":"2023-01-24T09:29:02Z","date_updated":"2023-01-24T09:29:02Z","creator":"dernst","checksum":"999e443b54e4fdaa2542ca5a97619731","relation":"main_file","access_level":"open_access","file_name":"2022_MolecularCell_Zapletal.pdf","content_type":"application/pdf","success":1,"file_id":"12354"}],"acknowledged_ssus":[{"_id":"EM-Fac"}],"abstract":[{"text":"MicroRNA (miRNA) and RNA interference (RNAi) pathways rely on small RNAs produced by Dicer endonucleases. Mammalian Dicer primarily supports the essential gene-regulating miRNA pathway, but how it is specifically adapted to miRNA biogenesis is unknown. We show that the adaptation entails a unique structural role of Dicer’s DExD/H helicase domain. Although mice tolerate loss of its putative ATPase function, the complete absence of the domain is lethal because it assures high-fidelity miRNA biogenesis. Structures of murine Dicer⋅miRNA precursor complexes revealed that the DExD/H domain has a helicase-unrelated structural function. It locks Dicer in a closed state, which facilitates miRNA precursor selection. Transition to a cleavage-competent open state is stimulated by Dicer-binding protein TARBP2. Absence of the DExD/H domain or its mutations unlocks the closed state, reduces substrate selectivity, and activates RNAi. Thus, the DExD/H domain structurally contributes to mammalian miRNA biogenesis and underlies mechanistical partitioning of miRNA and RNAi pathways.","lang":"eng"}],"acknowledgement":"We thank Kristian Vlahovicek (University of Zagreb) for support of bioinformatics analyses and Vladimir Benes (EMBL Sequencing Facility) and Genomics and Bioinformatics Core Facility at the Institute of Molecular Genetics for help with RNA sequencing. The main funding was provided by the Czech Science Foundation (EXPRO grant 20-03950X to P.S. and 22-19896S to R. Stefl). Early stages of the work were supported by European Research Council grants under the European Union’s Horizon 2020 Research and Innovation Programme (grants 647403 to P.S. and 649030 to R. Stefl). V.B., D.F.J., and F.H. were in part supported by PhD student fellowships from the Charles University; this work will be in part fulfilling requirements for a PhD degree as “school work.” Funding of D.Z. included the OP RDE project “Internal Grant Agency of Masaryk University” no. CZ.02.2.69/0.0/0.0/19_073/0016943. The Ministry of Education, Youth, and Sports of the Czech Republic (MEYS CR) provided institutional support for CEITEC 2020 project LQ1601. For technical support, we acknowledge EMBL Monterotondo’s genome engineering and transgenic core facilities, the Czech Centre for Phenogenomics at the Institute of Molecular Genetics (supported by RVO 68378050 from the Czech Academy of Sciences and LM2018126 and CZ.02.1.01/0.0/0.0/18_046/0015861 CCP Infrastructure Upgrade II from MEYS CR), the Cryo-EM and Proteomics Core Facilities (CEITEC, Masaryk University) supported by the CIISB research infrastructure (LM2018127 from MEYS CR), and support from the Scientific Service Units of ISTA through resources from the Electron Microscopy Facility. Computational resources included e-Infrastruktura CZ (LM2018140) and ELIXIR-CZ (LM2018131) projects by MEYS CR and the Croatian National Centres of Research Excellence in Personalized Healthcare (#KK.01.1.1.01.0010) and Data Science and Advanced Cooperative Systems (#KK.01.1.1.01.0009) projects funded by the European Structural and Investment Funds grants.","language":[{"iso":"eng"}],"pmid":1,"date_created":"2023-01-12T12:05:36Z","isi":1,"day":"03","has_accepted_license":"1","article_processing_charge":"No","publication_status":"published","quality_controlled":"1","date_updated":"2023-08-04T08:57:17Z","date_published":"2022-11-03T00:00:00Z","tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","short":"CC BY (4.0)","image":"/images/cc_by.png","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"},"publication_identifier":{"issn":["1097-2765"]},"intvolume":"        82","author":[{"full_name":"Zapletal, David","last_name":"Zapletal","first_name":"David"},{"full_name":"Taborska, Eliska","last_name":"Taborska","first_name":"Eliska"},{"full_name":"Pasulka, Josef","last_name":"Pasulka","first_name":"Josef"},{"last_name":"Malik","first_name":"Radek","full_name":"Malik, Radek"},{"full_name":"Kubicek, Karel","first_name":"Karel","last_name":"Kubicek"},{"last_name":"Zanova","first_name":"Martina","full_name":"Zanova, Martina"},{"last_name":"Much","first_name":"Christian","full_name":"Much, Christian"},{"last_name":"Sebesta","first_name":"Marek","full_name":"Sebesta, Marek"},{"full_name":"Buccheri, Valeria","last_name":"Buccheri","first_name":"Valeria"},{"full_name":"Horvat, Filip","last_name":"Horvat","first_name":"Filip"},{"full_name":"Jenickova, Irena","first_name":"Irena","last_name":"Jenickova"},{"last_name":"Prochazkova","first_name":"Michaela","full_name":"Prochazkova, Michaela"},{"full_name":"Prochazka, Jan","first_name":"Jan","last_name":"Prochazka"},{"last_name":"Pinkas","first_name":"Matyas","full_name":"Pinkas, Matyas"},{"last_name":"Novacek","first_name":"Jiri","full_name":"Novacek, Jiri"},{"last_name":"Joseph","first_name":"Diego F.","full_name":"Joseph, Diego F."},{"last_name":"Sedlacek","first_name":"Radislav","full_name":"Sedlacek, Radislav"},{"full_name":"Bernecky, Carrie A","orcid":"0000-0003-0893-7036","last_name":"Bernecky","id":"2CB9DFE2-F248-11E8-B48F-1D18A9856A87","first_name":"Carrie A"},{"full_name":"O’Carroll, Dónal","last_name":"O’Carroll","first_name":"Dónal"},{"full_name":"Stefl, Richard","last_name":"Stefl","first_name":"Richard"},{"first_name":"Petr","last_name":"Svoboda","full_name":"Svoboda, Petr"}],"keyword":["Cell Biology","Molecular Biology"],"type":"journal_article","oa_version":"Published Version","status":"public","publication":"Molecular Cell","ddc":["570"],"volume":82,"year":"2022","article_type":"original","scopus_import":"1","citation":{"mla":"Zapletal, David, et al. “Structural and Functional Basis of Mammalian MicroRNA Biogenesis by Dicer.” <i>Molecular Cell</i>, vol. 82, no. 21, Elsevier, 2022, p. 4064–4079.e13, doi:<a href=\"https://doi.org/10.1016/j.molcel.2022.10.010\">10.1016/j.molcel.2022.10.010</a>.","apa":"Zapletal, D., Taborska, E., Pasulka, J., Malik, R., Kubicek, K., Zanova, M., … Svoboda, P. (2022). Structural and functional basis of mammalian microRNA biogenesis by Dicer. <i>Molecular Cell</i>. Elsevier. <a href=\"https://doi.org/10.1016/j.molcel.2022.10.010\">https://doi.org/10.1016/j.molcel.2022.10.010</a>","short":"D. Zapletal, E. Taborska, J. Pasulka, R. Malik, K. Kubicek, M. Zanova, C. Much, M. Sebesta, V. Buccheri, F. Horvat, I. Jenickova, M. Prochazkova, J. Prochazka, M. Pinkas, J. Novacek, D.F. Joseph, R. Sedlacek, C. Bernecky, D. O’Carroll, R. Stefl, P. Svoboda, Molecular Cell 82 (2022) 4064–4079.e13.","chicago":"Zapletal, David, Eliska Taborska, Josef Pasulka, Radek Malik, Karel Kubicek, Martina Zanova, Christian Much, et al. “Structural and Functional Basis of Mammalian MicroRNA Biogenesis by Dicer.” <i>Molecular Cell</i>. Elsevier, 2022. <a href=\"https://doi.org/10.1016/j.molcel.2022.10.010\">https://doi.org/10.1016/j.molcel.2022.10.010</a>.","ieee":"D. Zapletal <i>et al.</i>, “Structural and functional basis of mammalian microRNA biogenesis by Dicer,” <i>Molecular Cell</i>, vol. 82, no. 21. Elsevier, p. 4064–4079.e13, 2022.","ama":"Zapletal D, Taborska E, Pasulka J, et al. Structural and functional basis of mammalian microRNA biogenesis by Dicer. <i>Molecular Cell</i>. 2022;82(21):4064-4079.e13. doi:<a href=\"https://doi.org/10.1016/j.molcel.2022.10.010\">10.1016/j.molcel.2022.10.010</a>","ista":"Zapletal D, Taborska E, Pasulka J, Malik R, Kubicek K, Zanova M, Much C, Sebesta M, Buccheri V, Horvat F, Jenickova I, Prochazkova M, Prochazka J, Pinkas M, Novacek J, Joseph DF, Sedlacek R, Bernecky C, O’Carroll D, Stefl R, Svoboda P. 2022. Structural and functional basis of mammalian microRNA biogenesis by Dicer. Molecular Cell. 82(21), 4064–4079.e13."},"_id":"12143","month":"11","page":"4064-4079.e13","department":[{"_id":"CaBe"}],"issue":"21","external_id":{"isi":["000898565300011"],"pmid":["36332606"]},"title":"Structural and functional basis of mammalian microRNA biogenesis by Dicer","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8"}