Spliceosome Assembly Pathways for Different Types of Alternative Splicing Converge during Commitment to Splice Site Pairing in the A Complex
| Autor: | Tara L. Crabb, Matthew V. Kotlajich, Klemens J. Hertel |
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| Rok vydání: | 2009 |
| Předmět: |
Spliceosome
Time Factors Computational biology Biology Adenosine Triphosphate Humans snRNP splice Molecular Biology U2 snRNP binding Genetics Splice site mutation Models Genetic Hydrolysis Alternative splicing Intron Exons Articles Cell Biology Introns Alternative Splicing Kinetics Multiprotein Complexes RNA splicing Spliceosomes RNA Splice Sites HeLa Cells |
| Zdroj: | Kotlajich, MV; Crabb, TL; & Hertel, KJ. (2009). Spliceosome assembly pathways for different types of alternative splicing converge during commitment to splice site pairing in the A complex. Molecular and Cellular Biology, 29(4), 1072-1082. doi: 10.1128/MCB.01071-08. UC Irvine: Retrieved from: http://www.escholarship.org/uc/item/4tw6z0t2 |
| ISSN: | 1098-5549 |
| Popis: | Differential splice site pairing establishes alternative splicing patterns resulting in the generation of multiple mRNA isoforms. This process is carried out by the spliceosome, which is activated by a series of sequential structural rearrangements of its five core snRNPs. To determine when splice sites become functionally paired, we carried out a series of kinetic trap experiments using pre-mRNAs that undergo alternative 5′ splice site selection or alternative exon inclusion. We show that commitment to splice site pairing in both cases occurs in the A complex, which is characterized by the ATP-dependent association of the U2 snRNP with the branch point. Interestingly, the timing of splice site pairing is independent of the intron or exon definition modes of splice site recognition. Using the ATP analog ATPγS, we showed that ATP hydrolysis is required for splice site pairing independent from U2 snRNP binding to the pre-mRNA. These results identify the A complex as the spliceosomal assembly step dedicated to splice site pairing and suggest that ATP hydrolysis locks splice sites into a splicing pattern after stable U2 snRNP association to the branch point. Copyright © 2009, American Society for Microbiology. All Rights Reserved. |
| Databáze: | OpenAIRE |
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