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Group II intron splicing in vivo by first-step hydrolysis

Nature volume 391, pages 915–918 (1998)Cite this article

Abstract

Group I, group II and spliceosomal introns splice by two sequential transesterification reactions1. For both spliceosomal and group II introns, the first-step reaction occurs by nucleophilic attack on the 5′ splice junction by the 2′ hydroxyl of an internal adenosine, forming a 2′–5′ phosphodiester branch in the intron. The second reaction joins the two exons with a 3′–5′ phosphodiester bond and releases intron lariat. In vitro, group II introns can self-splice by an efficient alternative pathway in which the first-step reaction occurs by hydrolysis. The resulting linear splicing intermediate participates in normal second-step reactions, forming spliced exon and linear intron RNAs2,3. Here we show that the group II intron first-step hydrolysis reaction occurs in vivo in place of transesterification in the mitochondria of yeast strains containing branch-site mutations. As expected, the mutations block branching, but surprisingly still allow accurate splicing. This hydrolysis pathway may have been a step in the evolution of splicing mechanisms.

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Figure 1: Group II intron splicing pathways and domain-6 mutants.
Figure 2: RNA blots showing splicing phenotypes of mutant and control strains.
Figure 3: Reverse-transcriptase primer extension analysis of IVS-E2 RNA.

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Acknowledgements

This work was supported by NIH grants to P.S.P. and A.M.P. M.P. was a fellow of the Robert A. Welch Foundation.

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Authors and Affiliations

  1. the Department of Molecular Biology and Oncology, University of Texas Southwestern Medical Center, Dallas, 75235-9148, Texas, USA

    Mircea Podar & Philip S. Perlman

  2. the Department of Biochemistry and Molecular Biophysics, Columbia University College of Physicians and Surgeons, New York, 10032, New York, USA

    Vi T. Chu & Anna Marie Pyle

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  1. Mircea Podar
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  2. Vi T. Chu
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  3. Anna Marie Pyle
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  4. Philip S. Perlman
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Correspondence to Philip S. Perlman.

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Podar, M., Chu, V., Pyle, A. et al. Group II intron splicing in vivo by first-step hydrolysis. Nature 391, 915–918 (1998). https://doi.org/10.1038/36142

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