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

k 2022

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

ZAPLETAL, David, Karel KUBÍČEK, Martina ZÁNOVÁ, Radek MALIK, Marek ŠEBESTA et. al.

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

Language

English

Type of outcome

Prezentace na konferencích

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

Czech Republic

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14310/22:00126100

Organization unit

Faculty of Science

Keywords in English

Dicer; RNA interference; RNAi; miRNA; siRNA

Změněno: 10/3/2023 13:38, Mgr. Pavla Foltynová, Ph.D.

Abstract

V originále

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

Citovat

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.

@proceedings{1862096,
   author = {Zapletal, David and Kubíček, Karel and Zánová, Martina and Malik, Radek and Šebesta, Marek and Nováček, Jiří and Svoboda, Petr and Štefl, Richard},
   booktitle = {Instruct Biennial Structural Biology ConferenceMuntgebouw, Utrecht, The Netherlands | 19-20 May, 2022},
   keywords = {Dicer; RNA interference; RNAi; miRNA; siRNA},
   language = {eng},
   title = {Cryo-EM reveals the structural basis for the regulatory function of HEL1 domain in vertebrate DICERs},
   year = {2022}
}

TY  - CONF
ID  - 1862096
AU  - Zapletal, David - Kubíček, Karel - Zánová, Martina - Malik, Radek - Šebesta, Marek - Nováček, Jiří - Svoboda, Petr - Štefl, Richard
PY  - 2022
TI  - Cryo-EM reveals the structural basis for the regulatory function of HEL1 domain in vertebrate DICERs
KW  - Dicer
KW  - RNA interference
KW  - RNAi
KW  - miRNA
KW  - siRNA
N2  - 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.
ER  -

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 \textit{Instruct Biennial Structural Biology ConferenceMuntgebouw, Utrecht, The Netherlands $|$ 19-20 May, 2022}. 2022.

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