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Mol Biol Cell. 2010 Nov 15;21(22):4020-7. doi: 10.1091/mbc.E10-08-0690. Epub 2010 Sep 29.

Highly ordered spatial organization of the structural long noncoding NEAT1 RNAs within paraspeckle nuclear bodies.

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  • 1Centre National de la Recherche Scientifique, Formation de Recherche en Evolution-3238, Institut André Lwoff, Villejuif 94800, France.

Abstract

Paraspeckles (PSPs) are nuclear bodies associated with the retention in the nucleus of specific mRNAs. Two isoforms of a long noncoding RNA (NEAT1_v1/Menε and NEAT1_v2/Menβ) are required for the integrity of PSPs. Here, we analyzed the molecular organization of PSPs by immuno- and in situ hybridization electron microscopy. Detection of the paraspeckle markers PSPC1 and P54NRB/NONO confirm the identity between PSPs and the previously described interchromatin granule-associated zones (IGAZs). High-resolution in situ hybridization of NEAT1 transcripts revealed a highly ordered organization of IGAZ/PSPs. Although the 3.7-kb NEAT1_v1 and the identical 5' end of the 22.7-kb NEAT1_v2 transcripts are confined to the periphery, central sequences of NEAT1_v2 are found within the electron-dense core of the bodies. Moreover, the 3' end of NEAT1_v2 also localize to the periphery, indicating possible architectures for IGAZ/PSPs. These results further suggest that the organization of NEAT1 transcripts constrains the geometry of these bodies. Accordingly, we observed in HeLa and NIH 3T3 cells that IGAZ/PSPs are elongated structures with a well-defined diameter. Our results provide new insight on the ability of noncoding RNAs to form subcellular structures.

PMID:
20881053
[PubMed - indexed for MEDLINE]
PMCID:
PMC2982136
Free PMC Article
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