Disruption of pre-mRNA splicing in vivo results in reorganization of splicing factors

O'Keefe, R. T., Mayeda, A., Sadowski, C. L., Krainer, A. R., Spector, D. L. (1994) Disruption of pre-mRNA splicing in vivo results in reorganization of splicing factors. Journal of Cell Biology, 124 (3). pp. 249-260. ISSN 0021-9525

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URL: http://www.ncbi.nlm.nih.gov/pubmed/8294510
DOI: 10.1083/jcb.124.3.249

Abstract

We have examined the functional significance of the organization of pre-mRNA splicing factors in a speckled distribution in the mammalian cell nucleus. Upon microinjection into living cells of oligonucleotides or antibodies that inhibit pre-mRNA splicing in vitro, we observed major changes in the organization of splicing factors in vivo. Interchromatin granule clusters became uniform in shape, decreased in number, and increased in both size and content of splicing factors, as measured by immunofluorescence. These changes were transient and the organization of splicing factors returned to their normal distribution by 24 h following microinjection. Microinjection of these oligonucleotides or antibodies also resulted in a reduction of transcription in vivo, but the oligonucleotides did not inhibit transcription in vitro. Control oligonucleotides did not disrupt splicing or transcription in vivo. We propose that the reorganization of splicing factors we observed is the result of the inhibition of splicing in vivo.

Item Type: Paper
Uncontrolled Keywords: Autoradiography Base Sequence Cell Nucleus HeLa Cells Humans Microinjections Microscopy, Electron Molecular Sequence Data Nuclear Proteins Oligodeoxyribonucleotides RNA Precursors RNA Splicing RNA, Small Nuclear Ribonucleoproteins Ribonucleoproteins, Small Nuclear Spliceosomes Transcription, Genetic
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > pre-mRNA
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > splicing factor
CSHL Authors:
Communities: CSHL labs > Krainer lab
CSHL labs > Spector lab
Depositing User: Matt Covey
Date Deposited: 10 Dec 2012 19:44
Last Modified: 29 Jan 2015 19:32
PMCID: PMC2119927
Related URLs:
URI: http://repository.cshl.edu/id/eprint/26329

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