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  Retention of posttranscriptional spliceosomes in nuclear speckles until splicing completion.

Girard, C., Will, C. L., Peng, J., Makarov, E. M., Kastner, B., Lemm, I., et al. (2012). Retention of posttranscriptional spliceosomes in nuclear speckles until splicing completion. Nature communications, 3: 994. doi:10.1038/ncomms1998.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-000F-A53B-9 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-3DA5-2
Genre: Journal Article

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Girard, C., Author
Will, C. L.1, Author              
Peng, J.1, Author              
Makarov, E. M.1, Author              
Kastner, B.1, Author              
Lemm, I.1, Author              
Urlaub, Henning2, Author              
Hartmuth, K.1, Author              
Lührmann, R.1, Author              
Affiliations:
1Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578576              
2Research Group of Bioanalytical Mass Spectrometry, MPI for biophysical chemistry, Max Planck Society, ou_578613              

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 Abstract: There is little quantitative information regarding how much splicing occurs co-transcriptionally in higher eukaryotes, and it remains unclear where precisely splicing occurs in the nucleus. Here we determine the global extent of co- and post-transcriptional splicing in mammalian cells, and their respective subnuclear locations, using antibodies that specifically recognize phosphorylated SF3b155 (P-SF3b155) found only in catalytically activated/active spliceosomes. Quantification of chromatin- and nucleoplasm-associated P-SF3b155 after fractionation of HeLa cell nuclei, reveals that ~80% of pre-mRNA splicing occurs co-transcriptionally. Active spliceosomes localize in situ to regions of decompacted chromatin, at the periphery of or within nuclear speckles. Immunofluorescence microscopy with anti-P-SF3b155 antibodies, coupled with transcription inhibition and a block in splicing after SF3b155 phosphorylation, indicates that post-transcriptional splicing occurs in nuclear speckles and that release of post-transcriptionally spliced mRNA from speckles is coupled to the nuclear mRNA export pathway. Our data provide new insights into when and where splicing occurs in cells.

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 Dates: 2012-08-07
 Publication Status: Published online
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 Rev. Method: Peer
 Identifiers: DOI: 10.1038/ncomms1998
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Title: Nature communications
Source Genre: Journal
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Pages: - Volume / Issue: 3 Sequence Number: 994 Start / End Page: - Identifier: -