title: NEAT1 long noncoding RNA regulates transcription via protein sequestration within subnuclear bodies.
abstract: Paraspeckles are subnuclear structures formed around nuclear paraspeckle assembly transcript 1 (NEAT1)/MENε/β long noncoding RNA (lncRNA). Here we show that paraspeckles become dramatically enlarged after proteasome inhibition. This enlargement is mainly caused by NEAT1 transcriptional up-regulation rather than accumulation of undegraded paraspeckle proteins. Of interest, however, using immuno-electron microscopy, we find that key paraspeckle proteins become effectively depleted from the nucleoplasm by 50% when paraspeckle assembly is enhanced, suggesting a sequestration mechanism. We also perform microarrays from NEAT1-knockdown cells and find that NEAT1 represses transcription of several genes, including the RNA-specific adenosine deaminase B2 (ADARB2) gene. In contrast, the NEAT1-binding paraspeckle protein splicing factor proline/glutamine-rich (SFPQ) is required for ADARB2 transcription. This leads us to hypothesize that ADARB2 expression is controlled by NEAT1-dependent sequestration of SFPQ. Accordingly, we find that ADARB2 expression is strongly reduced upon enhanced SFPQ sequestration by proteasome inhibition, with concomitant reduction in SFPQ binding to the ADARB2 promoter. Finally, NEAT1(-/-) fibroblasts are more sensitive to proteasome inhibition, which triggers cell death, suggesting that paraspeckles/NEAT1 attenuates the cell death pathway. These data further confirm that paraspeckles are stress-responsive nuclear bodies and provide a model in which induced NEAT1 controls target gene transcription by protein sequestration into paraspeckles.
authors.affiliation: Institute for Genetic Medicine, Hokkaido University, Sapporo 060-0815, Japan Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Tokyo 135-0064, Japan Centre National de la Recherche Scientifique, UMR-8122, Institut Gustave Roussy, Villejuif 94805, France Western Australian Institute for Medical Research, Center for Medical Research, University of Western Australia, Perth, Western Australia 6000, Australia Graduate School of Frontier Biosciences, Osaka University, Suita 565-0871, Japan Hitachi, Kokubunji, Tokyo 185-8601, Japan RIKEN Advanced Institute, Wako 351-0198, Japan Centre National de la Recherche Scientifique, FRE 3402, UPMC Université Paris 06, Paris 75 252, France.
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Hirose
authors.forename: Giorgio
authors.initials: G
authors.lastname: Virnicchi
authors.forename: Akie
authors.initials: A
authors.lastname: Tanigawa
authors.forename: Takao
authors.initials: T
authors.lastname: Naganuma
authors.forename: Ruohan
authors.initials: R
authors.lastname: Li
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kimura
authors.forename: Takahide
authors.initials: T
authors.lastname: Yokoi
authors.forename: Shinichi
authors.initials: S
authors.lastname: Nakagawa
authors.forename: Marianne
authors.initials: M
authors.lastname: Bénard
authors.forename: Archa H
authors.initials: AH
authors.lastname: Fox
authors.forename: Gérard
authors.initials: G
authors.lastname: Pierron
mesh_terms: D000243:Adenosine Deaminase / Q000235:genetics / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism* / Q000648:ultrastructure; D004268:DNA-Binding Proteins; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D007976:Leupeptins / Q000494:pharmacology; D034521:Nuclear Matrix-Associated Proteins / Q000378:metabolism; D050810:Octamer Transcription Factors / Q000378:metabolism; D011401:Promoter Regions, Genetic; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D061988:Proteasome Inhibitors / Q000494:pharmacology; D021381:Protein Transport; D059748:Proteolysis; D062085:RNA, Long Noncoding / Q000235:genetics* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D004268:DNA-Binding Proteins; D007976:Leupeptins; C577276:NEAT1 long non-coding RNA, human; C083214:NONO protein, human; D034521:Nuclear Matrix-Associated Proteins; D050810:Octamer Transcription Factors; D061988:Proteasome Inhibitors; D062085:RNA, Long Noncoding; D016601:RNA-Binding Proteins; D046988:Proteasome Endopeptidase Complex; C567955:ADARB1 protein, human; D000243:Adenosine Deaminase; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde