Many of the protein factors that play a role in nuclear export of mRNAs have been identified, but still little is known about how mRNAs are transported through the cell nucleus and which nuclear compartments are involved in mRNA transport. Using fluorescent 2'O-methyl oligoribonucleotide probes, we investigated the mobility of poly(A)+ RNA in the nucleoplasm and in nuclear speckles of U2OS cells. Quantitative analysis of diffusion using photobleaching techniques revealed that the majority of poly(A)+ RNA move throughout the nucleus, including in and out of speckles (also called SC-35 domains), which are enriched for splicing factors. Interestingly, in the presence of the transcription inhibitor 5,6-dichloro-1-β-d-ribofuranosylbenzimidazole, the association of poly(A)+ RNA with speckles remained dynamic. Our results show that RNA movement is energy dependent and that the proportion of nuclear poly(A)+ RNA that resides in speckles is a dynamic population that transiently interacts with speckles independent of the transcriptional status of the cell. Rather than the poly(A)+ RNA within speckles serving a stable structural role, our findings support the suggestion of a more active role of these regions in nuclear RNA metabolism and/or transport.
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26 April 2004
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April 26 2004
Poly(A) + RNAs roam the cell nucleus and pass through speckle domains in transcriptionally active and inactive cells
Chris Molenaar,
Chris Molenaar
Department of Molecular Cell Biology, Leiden University Medical Center, 2333 AL Leiden, Netherlands
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Abadir Abdulle,
Abadir Abdulle
Department of Molecular Cell Biology, Leiden University Medical Center, 2333 AL Leiden, Netherlands
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Aarti Gena,
Aarti Gena
Department of Molecular Cell Biology, Leiden University Medical Center, 2333 AL Leiden, Netherlands
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Hans J. Tanke,
Hans J. Tanke
Department of Molecular Cell Biology, Leiden University Medical Center, 2333 AL Leiden, Netherlands
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Roeland W. Dirks
Roeland W. Dirks
Department of Molecular Cell Biology, Leiden University Medical Center, 2333 AL Leiden, Netherlands
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Chris Molenaar
Department of Molecular Cell Biology, Leiden University Medical Center, 2333 AL Leiden, Netherlands
Abadir Abdulle
Department of Molecular Cell Biology, Leiden University Medical Center, 2333 AL Leiden, Netherlands
Aarti Gena
Department of Molecular Cell Biology, Leiden University Medical Center, 2333 AL Leiden, Netherlands
Hans J. Tanke
Department of Molecular Cell Biology, Leiden University Medical Center, 2333 AL Leiden, Netherlands
Roeland W. Dirks
Department of Molecular Cell Biology, Leiden University Medical Center, 2333 AL Leiden, Netherlands
Address correspondence to R.W. Dirks, Dept. of Molecular Cell Biology, Leiden University Medical Center, Wassenaarseweg 72, 2333 AL Leiden, Netherlands. Tel.: 31-71-5276026. Fax.: 31-71-5276180. email: [email protected]
Abbreviations list: DRB, 5,6-dichloro-1-β-d-ribofuranosylbenzimidazole; FLIP, fluorescence loss in photobleaching; HCMV, human cytomegalovirus; PABP2, poly(A) binding protein II; TAMRA, tetramethylrhodamine.
Received:
October 29 2003
Accepted:
March 22 2004
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2004
J Cell Biol (2004) 165 (2): 191–202.
Article history
Received:
October 29 2003
Accepted:
March 22 2004
Citation
Chris Molenaar, Abadir Abdulle, Aarti Gena, Hans J. Tanke, Roeland W. Dirks; Poly(A)+ RNAs roam the cell nucleus and pass through speckle domains in transcriptionally active and inactive cells . J Cell Biol 26 April 2004; 165 (2): 191–202. doi: https://doi.org/10.1083/jcb.200310139
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