Source:http://linkedlifedata.com/resource/pubmed/id/15194683
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
35
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| pubmed:dateCreated |
2004-8-23
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| pubmed:abstractText |
The mutually exclusive exons 2 and 3 of alpha-tropomyosin (alphaTM) have been used as a model system for strictly regulated alternative splicing. Exon 2 inclusion is only observed at high levels in smooth muscle (SM) tissues, whereas striated muscle and non-muscle cells use predominantly exon 3. Experiments in cell culture have shown that exon 2 selection results from repression of exon 3 and that this repression is mediated by regulatory elements flanking exon 3. We have now tested the cell culture-derived model in transgenic mice. We show that by harnessing the intronic splicing regulatory elements, expression of an enhanced green fluorescent protein transgene with a constitutively active promoter can be restricted to SM cells. Splicing of both endogenous alphaTM and a series of transgenes carrying regulatory element mutations was analyzed by reverse transcriptasePCR. These studies indicated that although SM-rich tissues are equipped to regulate splicing of high levels of endogenous or transgene alphaTM RNA, other non-SM tissues such as spleen, which express lower amounts of alphaTM, also splice significant proportions of exon 2, and this splicing pattern can be recapitulated by transgenes expressed at low levels. We confirm the importance in vivo of the negatively acting regulatory elements for regulated skipping of exon 3. Moreover, we provide evidence that some of the regulatory factors responsible for exon 3 skipping appear to be titratable, with loss of regulated splicing sometimes being associated with high transgene expression levels.
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| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Aug
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| pubmed:issn |
0021-9258
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:day |
27
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| pubmed:volume |
279
|
| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
36660-9
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| pubmed:dateRevised |
2008-11-21
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| pubmed:meshHeading |
pubmed-meshheading:15194683-Alternative Splicing,
pubmed-meshheading:15194683-Animals,
pubmed-meshheading:15194683-Base Sequence,
pubmed-meshheading:15194683-Exons,
pubmed-meshheading:15194683-Gene Expression Regulation,
pubmed-meshheading:15194683-Genetic Techniques,
pubmed-meshheading:15194683-Genetic Vectors,
pubmed-meshheading:15194683-Green Fluorescent Proteins,
pubmed-meshheading:15194683-Luminescent Proteins,
pubmed-meshheading:15194683-Mice,
pubmed-meshheading:15194683-Mice, Inbred C57BL,
pubmed-meshheading:15194683-Mice, Transgenic,
pubmed-meshheading:15194683-Microscopy, Fluorescence,
pubmed-meshheading:15194683-Models, Genetic,
pubmed-meshheading:15194683-Molecular Sequence Data,
pubmed-meshheading:15194683-Muscle, Smooth,
pubmed-meshheading:15194683-Mutation,
pubmed-meshheading:15194683-Plasmids,
pubmed-meshheading:15194683-Polymerase Chain Reaction,
pubmed-meshheading:15194683-Promoter Regions, Genetic,
pubmed-meshheading:15194683-RNA,
pubmed-meshheading:15194683-Reverse Transcriptase Polymerase Chain Reaction,
pubmed-meshheading:15194683-Spleen,
pubmed-meshheading:15194683-Tissue Distribution,
pubmed-meshheading:15194683-Transgenes,
pubmed-meshheading:15194683-Tropomyosin
|
| pubmed:year |
2004
|
| pubmed:articleTitle |
Regulated tissue-specific alternative splicing of enhanced green fluorescent protein transgenes conferred by alpha-tropomyosin regulatory elements in transgenic mice.
|
| pubmed:affiliation |
Department of Biochemistry, Tennis Court Road, University of Cambridge, Cambridge CB2 1QW, United Kingdom.
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|