Functional Domains of NEAT1 Architectural lncRNA Induce Paraspeckle Assembly through Phase Separation

Tomohiro Yamazaki, Sylvie Souquere, Takeshi Chujo, Simon Kobelke, Yee Seng Chong, Archa H. Fox, Charles S. Bond, Shinichi Nakagawa, Gerard Pierron, Tetsuro Hirose

Research output: Contribution to journalArticle

Abstract

A class of long noncoding RNAs (lncRNAs) has architectural functions in nuclear body construction; however, specific RNA domains dictating their architectural functions remain uninvestigated. Here, we identified the domains of the architectural NEAT1 lncRNA that construct paraspeckles. Systematic deletion of NEAT1 portions using CRISPR/Cas9 in haploid cells revealed modular domains of NEAT1 important for RNA stability, isoform switching, and paraspeckle assembly. The middle domain, containing functionally redundant subdomains, was responsible for paraspeckle assembly. Artificial tethering of the NONO protein to a NEAT1_2 mutant lacking the functional subdomains rescued paraspeckle assembly, and this required the NOPS dimerization domain of NONO. Paraspeckles exhibit phase-separated properties including susceptibility to 1,6-hexanediol treatment. RNA fragments of the NEAT1_2 subdomains preferentially bound NONO/SFPQ, leading to phase-separated aggregates in vitro. Thus, we demonstrate that the enrichment of NONO dimers on the redundant NEAT1_2 subdomains initiates construction of phase-separated paraspeckles, providing mechanistic insights into lncRNA-based nuclear body formation. The NEAT1 architectural noncoding RNA has a modular domain structure with distinct functions required for paraspeckle formation. Among the functional domains, the middle domain is composed of three distinct, functionally redundant, subdomains that recruit NONO dimers to initiate assembly of the phase-separated paraspeckle structure.

LanguageEnglish
Pages1038-1053.e7
JournalMolecular Cell
Volume701
Issue number6
DOIs
StatePublished - 21 Jun 2018

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Long Noncoding RNA
Clustered Regularly Interspaced Short Palindromic Repeats
RNA
RNA Isoforms
Untranslated RNA
Haploidy
RNA Stability
Dimerization
Proteins

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Yamazaki, Tomohiro ; Souquere, Sylvie ; Chujo, Takeshi ; Kobelke, Simon ; Chong, Yee Seng ; Fox, Archa H. ; Bond, Charles S. ; Nakagawa, Shinichi ; Pierron, Gerard ; Hirose, Tetsuro. / Functional Domains of NEAT1 Architectural lncRNA Induce Paraspeckle Assembly through Phase Separation. In: Molecular Cell. 2018 ; Vol. 701, No. 6. pp. 1038-1053.e7
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Yamazaki, T, Souquere, S, Chujo, T, Kobelke, S, Chong, YS, Fox, AH, Bond, CS, Nakagawa, S, Pierron, G & Hirose, T 2018, 'Functional Domains of NEAT1 Architectural lncRNA Induce Paraspeckle Assembly through Phase Separation' Molecular Cell, vol 701, no. 6, pp. 1038-1053.e7. DOI: 10.1016/j.molcel.2018.05.019

Functional Domains of NEAT1 Architectural lncRNA Induce Paraspeckle Assembly through Phase Separation. / Yamazaki, Tomohiro; Souquere, Sylvie; Chujo, Takeshi; Kobelke, Simon; Chong, Yee Seng; Fox, Archa H.; Bond, Charles S.; Nakagawa, Shinichi; Pierron, Gerard; Hirose, Tetsuro.

In: Molecular Cell, Vol. 701, No. 6, 21.06.2018, p. 1038-1053.e7.

Research output: Contribution to journalArticle

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AU - Yamazaki,Tomohiro

AU - Souquere,Sylvie

AU - Chujo,Takeshi

AU - Kobelke,Simon

AU - Chong,Yee Seng

AU - Fox,Archa H.

AU - Bond,Charles S.

AU - Nakagawa,Shinichi

AU - Pierron,Gerard

AU - Hirose,Tetsuro

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KW - paraspeckle

KW - phase separation

KW - prion-like domain

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