Abstract
Group II introns are self-splicing RNAs that are commonly found in the genes of plants, fungi, yeast and bacteria1,2. Little is known about the tertiary structure of group II introns, which are among the largest natural ribozymes. The most conserved region of the intron is domain 5 (D5), which, together with domain 1 (D1), is required for all reactions catalysed by the intron3. Despite the importance of D5, its spatial relationship and tertiary contacts to other active-site constituents have remained obscure. Furthermore, D5 has never been placed directly at a site of catalysis by the intron. Here we show that a set of tertiary interactions (λ–λ′) links catalytically essential regions of D5 and D1, creating the framework for an active-site and anchoring it at the 5′ splice site. Highly conserved elements similar to components of the λ–λ′ interaction are found in the eukaryotic spliceosome.
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Acknowledgements
We thank Q. Liu for preparing plasmids encoding exD123 mutants. We also thank S. Strobel for helpful discussions, R. Sousa for the clone of the Y639F RNA polymerase and P. S. Perlman for the gift of plasmids pJDI5′-75 and pJDI3′-673. M.B. is a Postdoctoral Research Associate and A.M.P. is an Assistant Investigator with the Howard Hughes Medical Institute, which we thank for financial support of this work. A.D.L. is supported by a fellowship from PRAXIS XXI (Portugal).
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Boudvillain, M., de Lencastre, A. & Pyle, A. A tertiary interaction that links active-site domains to the 5′ splice site of a group II intron. Nature 406, 315–318 (2000). https://doi.org/10.1038/35018589
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DOI: https://doi.org/10.1038/35018589
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