Regulation of Alternative Splicing in Tumor Metastasis
Annual rept. 1 Sep 1998-1 Sep 1999
TORONTO UNIV (ONTARIO)
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We have developed methods lot the identification end characterization of SR and SR-associated Wotcins with the aim of developing a more detailed understanding of interactions that are important for the regulation of pre-mRNA splicing. A splicing complex affinity-selection assay has been established that allows the sensitive detection of SR protein interactions at different stages of splicing complex formation. Using this assay, we have investigated the mechanism by which exonic splicing enhancer sequences ESEs function. in particular. we have tested a previous model in which SR proteins bound to an ESE promote the formation of splicing complexes at adjacent splice sites by stimulating the binding of the U2 snRNp auxiliary factor U2AF to the polypyrimidine tract The results clearly demonstrate that the recruitment of SR protein by an ESE to pre-mRNA does not influence the binding of the U2A-65kDa Subunit to the pre-mRNA. Moreover, evidence is obtained using the affinity selection assay that, instead of the ESE, the presence of Ul snRNP is critical for U2AF binding.
- Medicine and Medical Research