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Structural elucidation of a PRP8 core domain from the heart of the spliceosome

Nature Structural & Molecular Biology volume 15, pages 1199–1205 (2008)Cite this article

Abstract

The spliceosome is a complex ribonucleoprotein (RNP) particle containing five RNAs and more than 100 associated proteins. One of these proteins, PRP8, has been shown to interact directly with the splice sites and branch region of precursor-mRNAs (pre-mRNAs) and spliceosomal RNAs associated with catalysis of the two steps of splicing. The 1.85-Å X-ray structure of the core of PRP8 domain IV, implicated in key spliceosomal interactions, reveals a bipartite structure that includes the presence of an RNase H fold linked to a five-helix assembly. Analysis of mutant yeast alleles and cross-linking results in the context of this structure, coupled with RNA binding studies, suggests that domain IV forms a surface that interacts directly with the RNA structures at the catalytic core of the spliceosome.

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Figure 1: Structure of PRP8 domain IV.
Figure 2: Mapping of yeast mutant alleles and a 5′ splice site cross-link in the context of the PRP8 domain IV core.
Figure 3: Comparison of the PRP8 domain IV core with RNase H folds.
Figure 4: RNA Binding by PRP8 domain IV core.
Figure 5: Representations of the proposed PRP8 domain IV core RNA binding surface.

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Acknowledgements

We thank C. Guthrie (University of California, San Francisco) for providing yeast strain YTB72 and the plasmid pJU186MAS, B. Hazes for technical assistance, and M. Glover and T. Wu for helpful discussions. This work was supported by an Operating Grant from the Canadian Institutes of Health Research (CIHR) and by funding from the Alberta Synchrotron Institute.

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  1. Department of Biochemistry, 474 Medical Sciences Building, University of Alberta, Edmonton, T6G 2H7, Alberta, Canada

    Dustin B Ritchie, Matthew J Schellenberg, Emily M Gesner, Sheetal A Raithatha, David T Stuart & Andrew M MacMillan

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Correspondence to Andrew M MacMillan.

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Ritchie, D., Schellenberg, M., Gesner, E. et al. Structural elucidation of a PRP8 core domain from the heart of the spliceosome. Nat Struct Mol Biol 15, 1199–1205 (2008). https://doi.org/10.1038/nsmb.1505

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