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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3
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pubmed:dateCreated |
1996-1-4
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pubmed:abstractText |
We have devised an in vitro splicing assay in which the mutually exclusive exons 2 and 3 of alpha-tropomyosin act as competing 3' splice sites for joining to exon 1. Splicing in normal HeLa cell nuclear extracts results in almost exclusive joining of exons 1 and 3. Splicing in decreased nuclear extract concentrations and decreased ionic strength results in increased 1-2 splicing. We have used this assay to determine the role of three constitutive pre-mRNA splicing factors on alternative 3' splice site selection. Polypyrimidine tract binding protein (PTB) was found to inhibit the splicing of introns containing a strong binding site for this factor. However, the inhibitory effect of PTB could be partially reversed if pre-mRNAs were preincubated with U2 auxiliary factor (U2AF) prior to splicing in PTB-supplemented extracts. For alpha-tropomyosin, regulation of splicing by PTB and U2AF primarily affected the joining of exons 1-3 with no dramatic increases in 1-2 splicing being detected. Preincubation of pre-mRNAs with SR proteins led to small increases in 1-2 splicing. However, if pre-mRNAs were preincubated with SR proteins followed by splicing in PTB-supplemented extracts, there was a nearly complete reversal of the normal 1-2 to 1-3 splicing ratios. Thus, multiple pairwise, and sometimes antagonizing, interactions between constitutive pre-mRNA splicing factors and the pre-mRNA can regulate 3' splice site selection.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/DNA-Binding Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Nuclear Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphoproteins,
http://linkedlifedata.com/resource/pubmed/chemical/Polypyrimidine Tract-Binding Protein,
http://linkedlifedata.com/resource/pubmed/chemical/RNA-Binding Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Ribonucleoproteins,
http://linkedlifedata.com/resource/pubmed/chemical/Tropomyosin,
http://linkedlifedata.com/resource/pubmed/chemical/serine-arginine-rich splicing...,
http://linkedlifedata.com/resource/pubmed/chemical/splicing factor U2AF
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pubmed:status |
MEDLINE
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pubmed:month |
May
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pubmed:issn |
1355-8382
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
1
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
234-45
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pubmed:dateRevised |
2008-11-20
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pubmed:meshHeading |
pubmed-meshheading:7489496-Base Sequence,
pubmed-meshheading:7489496-Cell Nucleus,
pubmed-meshheading:7489496-DNA-Binding Proteins,
pubmed-meshheading:7489496-Exons,
pubmed-meshheading:7489496-Gene Expression Regulation,
pubmed-meshheading:7489496-HeLa Cells,
pubmed-meshheading:7489496-Humans,
pubmed-meshheading:7489496-Molecular Sequence Data,
pubmed-meshheading:7489496-Nuclear Proteins,
pubmed-meshheading:7489496-Phosphoproteins,
pubmed-meshheading:7489496-Polypyrimidine Tract-Binding Protein,
pubmed-meshheading:7489496-RNA Splicing,
pubmed-meshheading:7489496-RNA-Binding Proteins,
pubmed-meshheading:7489496-Ribonucleoproteins,
pubmed-meshheading:7489496-Subcellular Fractions,
pubmed-meshheading:7489496-Tropomyosin
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pubmed:year |
1995
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pubmed:articleTitle |
Regulation of alternative 3' splice site selection by constitutive splicing factors.
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pubmed:affiliation |
Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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