Factors Involved in the Activation of Pre-mRNA Splicing from Downstream Splicing Enhancers

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Abstract
The excision of introns with weak polypyrimidine tracts at their 3′ splice sites can be enhanced by sequence elements in the downstream exon or by a downstream 5′ splice site. The enhancers inside the exon do not conform to a strict consensus, but they are generally rich in purines. Here, we show that members of the family of SR proteins recognize these elements. Not only does SF2/ASF activate many different polypurine enhancers, but also at least one other SR protein, most likely SC35, is active as well. The degree of splicing activation varies with the polypurine enhancers and the SR proteins. Further, we show that the similar activation by downstream 5′ splice sites requires U1 snRNP, which is not the case with purine-rich enhancers. These results are consistent with a model showing that U1 snRNP binds to the 5′ splice site and SR proteins to exonic sequences upstream of the 5′ splice site. Both interact with U2AF at the 3′ splice site. This represents a molecular explanation for the exon recognition which is important for splice site selection in mammals.

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