In vivo analysis of the stability and transport of nuclear poly(A)+ RNA

Huang, S., Deerinck, T. J., Ellisman, M. H. , Spector, D. L. (August 1994) In vivo analysis of the stability and transport of nuclear poly(A)+ RNA. Journal of Cell Biology, 126 (4). pp. 877-99. ISSN 0021-9525

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DOI: 10.1083/jcb.126.4.877


We have studied the distribution of poly(A)+ RNA in the mammalian cell nucleus and its transport through nuclear pores by fluorescence and electron microscopic in situ hybridization. Poly(A)+ RNA was detected in the nucleus as a speckled pattern which includes interchromatin granule clusters and perichromatin fibrils. When cells are fractionated by detergent and salt extraction as well as DNase I digestion, the majority of the nuclear poly(A)+ RNA was found to remain associated with the nonchromatin RNP-enriched fraction of the nucleus. After inhibition of RNA polymerase II transcription for 5-10 h, a stable population of poly(A)+ RNA remained in the nucleus and was reorganized into fewer and larger interchromatin granule clusters along with pre-mRNA splicing factors. This stable population of nuclear RNA may play an important role in nuclear function. Furthermore, we have observed that, in actively transcribing cells, the regions of poly(A)+ RNA which reached the nuclear pore complexes appeared as narrow concentrations of RNA suggesting a limited or directed pathway of movement. All of the observed nuclear pores contained poly(A)+ RNA staining suggesting that they are all capable of exporting RNA. In addition, we have directly visualized, for the first time in mammalian cells, the transport of poly(A)+ RNA through the nuclear pore complexes.

Item Type: Paper
Uncontrolled Keywords: Animals CHO Cells Cell Line Cell Nucleus metabolism ultrastructure Chromatin ultrastructure Cricetinae Cytoplasmic Granules ultrastructure Hela Cells Humans In Situ Hybridization Microscopy Electron Microscopy Fluorescence Nuclear Matrix metabolism ultrastructure Oligodeoxyribonucleotides Oligonucleotide Probes Poly A analysis metabolism RNA analysis metabolism RNA Messenger
Subjects: organism description > animal > mammal
organs, tissues, organelles, cell types and functions > organelles, types and functions > nucleus
physics > biophysics > pore dynamics
organs, tissues, organelles, cell types and functions > tissues types and functions > transport > RNA transport
CSHL Authors:
Communities: CSHL labs > Spector lab
Depositing User: Brian Soldo
Date: August 1994
Date Deposited: 11 Apr 2012 18:06
Last Modified: 19 Mar 2013 19:59
PMCID: PMC2120126
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