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The RNA splicing factor hSlu7 is required for correct 3′ splice-site choice

Abstract

The production of correctly spliced messenger RNA requires two catalytic splicing steps1,2,3. During step II, exon 1 attacks an adenine-guanine (AG) dinucleotide at the 3′ splice site. This AG is usually located between 18 and 40 nucleotides downstream from the branch site, and closer AGs are skipped in favour of AGs located more optimally downstream4,5,6. At present, little is understood about how the correct AG is distinguished from other AGs. Here we describe a metazoan splicing factor (hSlu7) that is required for selection of the correct AG. In the absence of hSlu7, use of the correct AG is suppressed and incorrect AGs are activated. We investigated this loss of fidelity by analysing spliceosomes assembled in the absence of hSlu7. These studies reveal that exon 1 is loosely associated with these spliceosomes. Thus, the improperly held exon cannot access the correct AG, but can attack other AGs indiscriminately. We conclude that hSlu7 is required to hold exon 1 tightly within the spliceosome for attack on a prespecified AG.

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Figure 1: Suppression of the correct AG and activation of aberrant upstream AGs in the absence of hSlu7.
Figure 2: Suppression of the correct AG and activation of aberrant downstream AGs in the absence of hSlu7.
Figure 3: The attacking exon is not properly associated with ΔhSlu7 spliceosomes.
Figure 4: The improperly held exon 1 attacks aberrant AGs.

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Acknowledgements

We thank K. Clouse, O. Gozani, R. Gaur, B. Graveley, K. Hertel and Z. Zhou for critical comments on the manuscript. This work was supported by an NIH grant to R.R.

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Correspondence to Robin Reed.

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Chua, K., Reed, R. The RNA splicing factor hSlu7 is required for correct 3′ splice-site choice. Nature 402, 207–210 (1999). https://doi.org/10.1038/46086

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