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PMID: 1532583 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Transcription-dependent colocalization of the U1, U2, U4/U6, and U5 snRNPs in coiled bodies.

The Journal of cell biology ·Vol. 117 ·No. 1 ·1992-04-00 ·Pages 1-14

Carmo-Fonseca M, Pepperkok R, Carvalho MT, Lamond AI

Abstract

We have recently shown that discrete foci are present in the nuclei of mammalian cells in which each of the U1, U2, U4/U6, and U5 snRNPs involved in pre-mRNA splicing, and the non-snRNP-splicing factor U2AF, are concentrated (Carmo-Fonseca, M., D. Tollervey, R. Pepperkok, S. Barabino, A. Merdes, C. Brunner, P. D. Zamore, M. R. Green, E. Hurt, and A. I. Lamond. 1991. EMBO (Eur. Mol. Biol. Organ.) J. 10:195-206; Carmo-Fonseca, M., R. Pepperkok, B. S. Sproat, W. Ansorge, M. S. Swanson, and A. I. Lamond. 1991 EMBO (Eur. Mol. Biol. Organ.) J. 10:1863-1873). Here, we identify these snRNP-rich organelles as coiled bodies. snRNPs no longer concentrate in coiled bodies after cells are treated with the transcription inhibitors alpha-amanitin or actinomycin D. snRNP association with coiled bodies is also disrupted by heat shock. This indicates that the association of snRNPs with coiled bodies may be connected with the metabolism of nascent transcripts. A novel labeling method is described which shows both the RNA and protein components of individual snRNPs colocalizing in situ. Using this procedure all spliceosomal snRNPs are seen distributed in a nonhomogeneous pattern throughout the nucleoplasm, excluding nucleoli. They are most concentrated in coiled bodies, but in addition are present in "speckled" structures which are distinct from coiled bodies and which contain the non-snRNP splicing factor SC-35. U1 snRNP shows a more widespread nucleoplasmic staining, outside of coiled bodies and "speckled" structures, relative to the other snRNPs. The association of snRNPs with "speckles" is disrupted by heat shock but enhanced when cells are treated with alpha-amanitin.

MeSH Terms
Amanitins/pharmacology Antisense Elements (Genetics) Base Sequence Cell Nucleus/physiology,ultrastructure Dactinomycin/pharmacology HeLa Cells Humans Molecular Sequence Data Oligodeoxyribonucleotides Organelles/drug effects,physiology,ultrastructure RNA, Small Nuclear/genetics Ribonucleoproteins/analysis,genetics Ribonucleoproteins, Small Nuclear Transcription, Genetic/drug effects
Chemicals
Amanitins Antisense Elements (Genetics) Oligodeoxyribonucleotides RNA, Small Nuclear Ribonucleoproteins Ribonucleoproteins, Small Nuclear Dactinomycin
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Carmo-Fonseca M
European Molecular Biology Laboratory, Heidelberg, Germany.
Pepperkok R
Carvalho M T
Lamond A I
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.117.1.1
Published
1992-04-00
Pages
1-14
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289407
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.117.1.1 PMC Full Text View journal details
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