Recognition of the 5' splice site by the spliceosome.
The splicing of nuclear pre-mRNAs is catalyzed by a large, multicomponent ribonucleoprotein complex termed the spliceosome. Elucidation of the molecular mechanism of splicing identified small nuclear RNAs (snRNAs) as important components of the spliceosome, which, by analogy to the self-splicing group II introns, are implicated in formation of the catalytic center. In particular, the 5' splice site (5'SS) and the branch site, which represent the two substrates for the first step of splicing, are first recognized by U1 and U2 snRNPs, respectively. This initial recognition of splice sites is responsible for the global definition of exons and introns, and represents the primary target for regulation of splicing. Subsequently, pairing interaction between the 5'SS and U1 snRNA is disrupted and replaced by a new interaction of the 5'SS with U6 snRNA. The 5'SS signal contains an invariant GU dinucleotide present at the 5' end of nearly all known introns, however, the mechanism by which the spliceosome recognizes this element is not known. We have identified and characterized a specific UV light-induced crosslink formed between the 5'SS RNA and hPrp8, a protein component of U5 snRNP in the spliceosome that is likely to reflect a specific recognition of the GU dinucleotide for splicing. Because recognition of the 5'SS must be linked to formation of the catalytic site, the identification of a specific and direct interaction between the 5'SS and Prp8 has significant implications for the role of this protein in the mechanism of mRNA splicing.
The Rockefeller University, New York, NY 10021, USA. konarsk@rockvax.rockefeller.edu
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D002352:Carrier Proteins / Q000235:genetics; D005091:Exons; D006801:Humans; D007438:Introns; D008957:Models, Genetic; D012326:RNA Splicing; D016601:RNA-Binding Proteins; D017415:Ribonucleoprotein, U5 Small Nuclear / Q000235:genetics; D017381:Spliceosomes / Q000235:genetics* / Q000502:physiology*
D002352:Carrier Proteins; C115374:PRPF8 protein, human; D016601:RNA-Binding Proteins; D017415:Ribonucleoprotein, U5 Small Nuclear