Rapid, diffusional shuttling of poly(A) RNA between nuclear speckles and the nucleoplasm

Politz, J. C. R., Tuft, R. A., Prasanth, K. V. , Baudendistel, N., Fogarty, K. E., Lifshitz, L. M., Langowski, J., Spector, D. L., Pederson, T. (2006) Rapid, diffusional shuttling of poly(A) RNA between nuclear speckles and the nucleoplasm. Molecular Biology of the Cell, 17 (3). pp. 1239-1249. ISSN 10591524

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URL: http://www.ncbi.nlm.nih.gov/pubmed/16371503
DOI: 10.1091/mbc.E05-10-0952

Abstract

Speckles are nuclear bodies that contain pre-mRNA splicing factors and polyadenylated RNA. Because nuclear poly(A) RNA consists of both mRNA transcripts and nucleus-restricted RNAs, we tested whether poly(A) RNA in speckles is dynamic or rather an immobile, perhaps structural, component. Fluorescein-labeled oligo(dT) was introduced into HeLa cells stably expressing a red fluorescent protein chimera of the splicing factor SC35 and allowed to hybridize. Fluorescence correlation spectroscopy (FCS) showed that the mobility of the tagged poly(A) RNA was virtually identical in both speckles and at random nucleoplasmic sites. This same result was observed in photoactivation-tracking studies in which caged fluorescein-labeled oligo(dT) was used as hybridization probe, and the rate of movement away from either a speckle or nucleoplasmic site was monitored using digital imaging microscopy after photoactivation. Furthermore, the tagged poly(A) RNA was observed to rapidly distribute throughout the entire nucleoplasm and other speckles, regardless of whether the tracking observations were initiated in a speckle or the nucleoplasm. Finally, in both FCS and photoactivation-tracking studies, a temperature reduction from 37 to 22°C had no discernible effect on the behavior of poly(A) RNA in either speckles or the nucleoplasm, strongly suggesting that its movement in and out of speckles does not require metabolic energy. © 2006 by The American Society for Cell Biology.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > nuclear speckles
CSHL Authors:
Communities: CSHL labs > Spector lab
Depositing User: CSHL Librarian
Date Deposited: 09 Dec 2011 19:30
Last Modified: 28 Jan 2015 21:27
Related URLs:
URI: http://repository.cshl.edu/id/eprint/22878

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