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.

Základní údaje

Originální název

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

Název česky

Cryo-EM odhaluje strukturní podstatu regulační funkce HEL1 domény DICERu v obratlovcích

Autoři

ZAPLETAL, David (203 Česká republika, domácí), Karel KUBÍČEK (203 Česká republika, domácí), Martina ZÁNOVÁ (703 Slovensko), Radek MALIK, Marek ŠEBESTA (703 Slovensko, domácí), Jiří NOVÁČEK (203 Česká republika, domácí), Petr SVOBODA (203 Česká republika) a Richard ŠTEFL (203 Česká republika, garant, domácí)

Vydání

Instruct Biennial Structural Biology ConferenceMuntgebouw, Utrecht, The Netherlands | 19-20 May, 2022, 2022

Další údaje

Jazyk

angličtina

Typ výsledku

Prezentace na konferencích

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Kód RIV

RIV/00216224:14310/22:00126100

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

Dicer; RNA interference; RNAi; miRNA; siRNA

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

Anotace

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.

Návaznosti

EF19_073/0016943, projekt VaV

Název: Interní grantová agentura Masarykovy univerzity

Citovat

ZAPLETAL, David, Karel KUBÍČEK, Martina ZÁNOVÁ, Radek MALIK, Marek ŠEBESTA, Jiří NOVÁČEK, Petr SVOBODA a 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 a 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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