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Group II intron splicing in vivo by first-step hydrolysis.

Authors
  • Podar, M
  • Chu, V T
  • Pyle, A M
  • Perlman, P S
Type
Published Article
Journal
Nature
Publication Date
Feb 26, 1998
Volume
391
Issue
6670
Pages
915–918
Identifiers
PMID: 9495347
Source
Medline
License
Unknown

Abstract

Group I, group II and spliceosomal introns splice by two sequential transesterification reactions. 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 RNAs. 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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