Detailed Information on Publication Record

ZAPLETAL, David, Karel KUBÍČEK, Martina ZÁNOVÁ, Radek MALIK, Marek ŠEBESTA, Jiří NOVÁČEK, Petr SVOBODA and Richard ŠTEFL. Cryo-EM reveals the structural basis for the regulatory function of HEL1 domain in vertebrate DICERs. In Instruct Biennial Structural Biology ConferenceMuntgebouw, Utrecht, The Netherlands | 19-20 May, 2022. 2022.

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Basic information
Original name	Cryo-EM reveals the structural basis for the regulatory function of HEL1 domain in vertebrate DICERs
Name in Czech	Cryo-EM odhaluje strukturní podstatu regulační funkce HEL1 domény DICERu v obratlovcích
Authors	ZAPLETAL, David (203 Czech Republic, belonging to the institution), Karel KUBÍČEK (203 Czech Republic, belonging to the institution), Martina ZÁNOVÁ (703 Slovakia), Radek MALIK, Marek ŠEBESTA (703 Slovakia, belonging to the institution), Jiří NOVÁČEK (203 Czech Republic, belonging to the institution), Petr SVOBODA (203 Czech Republic) and Richard ŠTEFL (203 Czech Republic, guarantor, belonging to the institution).
Edition	Instruct Biennial Structural Biology ConferenceMuntgebouw, Utrecht, The Netherlands \| 19-20 May, 2022, 2022.

Other information
Original language	English
Type of outcome	Presentations at conferences
Field of Study	10608 Biochemistry and molecular biology
Country of publisher	Czech Republic
Confidentiality degree	is not subject to a state or trade secret
RIV identification code	RIV/00216224:14310/22:00126100
Organization unit	Faculty of Science
Keywords in English	Dicer; RNA interference; RNAi; miRNA; siRNA
Changed by	Changed by: Mgr. Pavla Foltynová, Ph.D., učo 106624. Changed: 10/3/2023 13:38.

Abstract
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.

Abstract

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.

Links
EF19_073/0016943, research and development project	Name: Interní grantová agentura Masarykovy univerzity