An architectural role for a nuclear noncoding RNA: NEAT1 RNA is essential for the structure of paraspeckles.
NEAT1 RNA, a highly abundant 4 kb ncRNA, is retained in nuclei in approximately 10 to 20 large foci that we show are completely coincident with paraspeckles, nuclear domains implicated in mRNA nuclear retention. Depletion of NEAT1 RNA via RNAi eradicates paraspeckles, suggesting that it controls sequestration of the paraspeckle proteins PSP1 and p54, factors linked to A-I editing. Unlike overexpression of PSP1, NEAT1 overexpression increases paraspeckle number, and paraspeckles emanate exclusively from the NEAT1 transcription site. The PSP-1 RNA binding domain is required for its colocalization with NEAT1 RNA in paraspeckles, and biochemical analyses support that NEAT1 RNA binds with paraspeckle proteins. Unlike other nuclear-retained RNAs, NEAT1 RNA is not A-I edited, consistent with a structural role in paraspeckles. Collectively, results demonstrate that NEAT1 functions as an essential structural determinant of paraspeckles, providing a precedent for a ncRNA as the foundation of a nuclear domain.
Clemson
Christine M
CM
University of Massachusetts Medical Center, 55 Lake Avenue North, Worcester, MA 01655, USA.
Hutchinson
John N
JN
Sara
Sergio A
SA
Ensminger
Alexander W
AW
Fox
Archa H
AH
Chess
Andrew
A
Lawrence
Jeanne B
JB
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D060365:Chloroplast Proteins; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006801:Humans; D047468:Immunoprecipitation; D045586:Intranuclear Inclusion Bodies / Q000737:chemistry*; D051379:Mice; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism*; D034622:RNA Interference; D012342:RNA, Small Nuclear / Q000502:physiology*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism
D060365:Chloroplast Proteins; D009687:Nuclear Proteins; C450059:PSPC1 protein, human; D012342:RNA, Small Nuclear; D016601:RNA-Binding Proteins; C494389:paraspeckle protein 1, mouse; D004722:Endoribonucleases; C106848:chloroplast RNA-binding protein p54