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Influence of Polymerase II Processivity on Alternative Splicing Depends on Splice Site Strength
- Source :
- Journal of Biological Chemistry. 278:52166-52171
- Publication Year :
- 2003
- Publisher :
- Elsevier BV, 2003.
-
Abstract
- Transcription and pre-mRNA splicing are coordinated temporally and spatially, and both processes can influence each other. In particular, control of transcriptional elongation by RNA polymerase II has proved to be important for alternative splicing regulation. In this report we demonstrate that the efficiency of exon recognition by the splicing machinery is crucial for the elongation control. Alternative splicing of the fibronectin extra domain I (EDI) is because the polypyrimidine tract of its 3'-splice site occurs suboptimal. By mutating the polypyrimidine tract of EDI in two different positions, individually or in combination, and by disrupting its exonic splicing silencer, we managed to generate minigenes with increasing degrees of exon recognition. Improvement of exon recognition is evidenced by independence from the splicing regulator SF2/ASF for inclusion. The mutated minigenes were used to transfect human cells in culture and study the responsiveness of EDI alternative splicing to activation or inhibition of pol II elongation. Our results revealed that responsiveness of exon skipping to elongation is inversely proportional to 3'-splice site strength, which means that the better the alternative exon is recognized by the splicing machinery, the less its degree of inclusion is affected by transcriptional elongation.
- Subjects :
- Time Factors
Transcription, Genetic
RNA Splicing
Exonic splicing enhancer
Biology
Transfection
Models, Biological
Biochemistry
Cell Line
Exon
Cell Line, Tumor
Humans
Point Mutation
RNA, Messenger
Phosphorylation
Molecular Biology
Exonic splicing silencer
Genetics
Binding Sites
Polymorphism, Genetic
Alternative splicing
Herpes Simplex Virus Protein Vmw65
Exons
Cell Biology
Exon skipping
Protein Structure, Tertiary
Alternative Splicing
Pyrimidines
Polypyrimidine tract
Mutation
RNA splicing
RNA Polymerase II
Plasmids
Minigene
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 278
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi.dedup.....e1545d61554248266a2da0d5e8449783