Cryo-EM reveals the structural basis for the regulatory function of HEL1 domain in vertebrate DICERs

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F22%3A00126100" target="_blank" >RIV/00216224:14310/22:00126100 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Cryo-EM reveals the structural basis for the regulatory function of HEL1 domain in vertebrate DICERs

Popis výsledku v původním jazyce

Dicer is RNase III enzyme producing small RNA in RNA silencing pathways. In mice, there are two isoforms that differ by presence or absence of the N-terminal HEL1 domain. The full-length Dicer acts in microRNA (miRNA) biogenesis while the truncated version, present in oocytes (DicerO), generates small interfering RNAs (siRNA) in RNA interference (RNAi) pathway. While models for siRNA and miRNA processing by vertebrate Dicer have emerged from the structural and biochemical studies, the active dicing state in Dicer-RNA structures has not been structurally characterized yet. In this study we used cryo-electron microscopy, AI-based prediction, and biochemical approaches to understand the structural difference between Dicer and DicerO, explaining their different in vivo functions. We observed that Dicer-RNA complex exists prevalently in an inactive, pre-dicing state (Fig.1b), while DicerO-RNA was observed almost exclusively in dicing-state (Fig.2c), revealing the molecular principles of substrate selection and enzymatic activity. Our data suggest HEL1 role in Dicer activity and substrate specificity by regulation of the loading of RNA substrate.

Název v anglickém jazyce

Cryo-EM reveals the structural basis for the regulatory function of HEL1 domain in vertebrate DICERs

Popis výsledku anglicky

Dicer is RNase III enzyme producing small RNA in RNA silencing pathways. In mice, there are two isoforms that differ by presence or absence of the N-terminal HEL1 domain. The full-length Dicer acts in microRNA (miRNA) biogenesis while the truncated version, present in oocytes (DicerO), generates small interfering RNAs (siRNA) in RNA interference (RNAi) pathway. While models for siRNA and miRNA processing by vertebrate Dicer have emerged from the structural and biochemical studies, the active dicing state in Dicer-RNA structures has not been structurally characterized yet. In this study we used cryo-electron microscopy, AI-based prediction, and biochemical approaches to understand the structural difference between Dicer and DicerO, explaining their different in vivo functions. We observed that Dicer-RNA complex exists prevalently in an inactive, pre-dicing state (Fig.1b), while DicerO-RNA was observed almost exclusively in dicing-state (Fig.2c), revealing the molecular principles of substrate selection and enzymatic activity. Our data suggest HEL1 role in Dicer activity and substrate specificity by regulation of the loading of RNA substrate.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/EF19_073%2F0016943" target="_blank" >EF19_073/0016943: Interní grantová agentura Masarykovy univerzity</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>S - Specificky vyzkum na vysokych skolach<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2022

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů