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PMID: 19536130 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Exon, intron and splice site locations in the spliceosomal B complex.

The EMBO journal ·Vol. 28 ·No. 15 ·2009-08-05 ·Pages 2283-92

Wolf E, Kastner B, Deckert J, Merz C, Stark H, Lührmann R

Abstract

In recent years, electron microscopy (EM) has allowed the generation of three-dimensional structure maps of several spliceosomal complexes. However, owing to their limited resolution, little is known at present about the location of the pre-mRNA, the spliceosomal small nuclear ribonucleoprotein or the spliceosome's active site within these structures. In this work, we used EM to localise the intron and the 5' and 3' exons of a model pre-mRNA, as well as the U2-associated protein SF3b155, in pre-catalytic spliceosomes (i.e. B complexes) by labelling them with an antibody that bears colloidal gold. Our data reveal that the intron and both exons, together with SF3b155, are located in specific regions of the head domain of the B complex. These results represent an important first step towards identifying functional sites in the spliceosome. The gold-labelling method adopted here can be applied to other spliceosomal complexes and may thus contribute significantly to our overall understanding of the pre-mRNA splicing process.

MeSH Terms
Exons Gold HeLa Cells Humans Introns Microscopy, Electron, Transmission Phosphoproteins/analysis RNA Precursors/analysis RNA Splice Sites RNA Splicing Factors Ribonucleoprotein, U2 Small Nuclear/analysis Spliceosomes/chemistry,ultrastructure Staining and Labeling/methods
Chemicals
Phosphoproteins RNA Precursors RNA Splice Sites RNA Splicing Factors Ribonucleoprotein, U2 Small Nuclear SF3B1 protein, human Gold
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Wolf Elmar
Department of Cellular Biochemistry, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
Kastner Berthold
Deckert Jochen
Merz Christian
Stark Holger
Lührmann Reinhard
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
1460-2075
DOI
10.1038/emboj.2009.171
Published
2009-08-05
Epub
2009-00-18
Pages
2283-92
Language
English
Region
England
NLM ID
8208664
PMCID
PMC2726701
Subset
IM
External Links
PubMed source DOI: 10.1038/emboj.2009.171 PMC Full Text View journal details
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