Abstract
The Down syndrome cell adhesion molecule (Dscam) gene has essential roles in neural wiring and pathogen recognition in Drosophila melanogaster. Dscam encodes 38,016 distinct isoforms via extensive alternative splicing. The 95 alternative exons in Dscam are organized into clusters that are spliced in a mutually exclusive manner. The exon 6 cluster contains 48 variable exons and uses a complex system of competing RNA structures to ensure that only one variable exon is included. Here we show that the heterogeneous nuclear ribonucleoprotein hrp36 acts specifically within, and throughout, the exon 6 cluster to prevent the inclusion of multiple exons. Moreover, hrp36 prevents serine/arginine-rich proteins from promoting the ectopic inclusion of multiple exon 6 variants. Thus, the fidelity of mutually exclusive splicing in the exon 6 cluster is governed by an intricate combination of alternative RNA structures and a globally acting splicing repressor.
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Acknowledgements
We thank members of the Graveley lab for discussions and comments on the manuscript. This work was funded by the Crick-Jacobs Center for Theoretical and Computational Biology (G.W.Y.), a long-term fellowship from the Human Frontier Science Program to M.B., and grants from the US National Institutes of Health to D.C.R. (GM61987) and B.R.G. (GM62561 and GM67842).
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S.O., M.B., J.P., Y.S. and J.M.Y. performed the experiments; S.O., M.B., J.P., Y.S., J.M.Y., D.C.R. and B.R.G. designed the experiments; S.O., M.B., J.P., Y.S., J.M.Y., G.W.Y., D.C.R. and B.R.G. analyzed the data; B.R.G. wrote the paper with input from all authors.
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J.M.Y. is employed by Illumina, Inc. She also performed assays and provided reagents for the DASL assays.
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Olson, S., Blanchette, M., Park, J. et al. A regulator of Dscam mutually exclusive splicing fidelity. Nat Struct Mol Biol 14, 1134–1140 (2007). https://doi.org/10.1038/nsmb1339
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DOI: https://doi.org/10.1038/nsmb1339
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