title: The Cbp2 protein suppresses splice site mutations in a group I intron.
abstract: The Cbp2 protein facilitates the folding of a group I intron in the COB pre-mRNA of yeast mitochondria. Based on its ability to suppress mutations affecting the auto-catalytic reaction, the protein appears to play a role in the selection of splice sites. Adding Cbp2 did not overcome the effects of mutations in P1 whose primary effect was on the first step of splicing. In contrast, most mutations affecting the ligation of exons were suppressed in vitro by Cbp2. These included mutations in P1, P9.0 and P10. In fact, a mutant transcript lacking both P9.0 and P10 ligated efficiently in the presence of Cbp2. P9.0 and P10 mutations also reduced the rate of cleavage at the 5' splice junction, and this effect was only partially mitigated by adding Cbp2. A competitive secondary structure near the 3' splice junction blocked Cbp2-stimulated splicing, but this mutation could be suppressed by co-transcriptional splicing in the presence of Cbp2. Our data underscore the importance of the interaction between the 5' and 3' splice junctions in group I introns and suggest that nucleotide-nucleotide interactions that stabilize the structure of group I introns can be superceded by protein-RNA interactions.
authors.affiliation: Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Gainesville 32610-0266, USA.
authors.forename: L C
authors.initials: LC
authors.lastname: Shaw
authors.forename: J
authors.initials: J
authors.lastname: Thomas
authors.forename: A S
authors.initials: AS
authors.lastname: Lewin
mesh_terms: D001059:Apoproteins / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D002467:Cell Nucleus / Q000235:genetics; D003573:Cytochrome b Group / Q000235:genetics; D045303:Cytochromes b; D005091:Exons; D005656:Fungal Proteins / Q000378:metabolism*; D007438:Introns / Q000235:genetics*; D008928:Mitochondria / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D012326:RNA Splicing / Q000235:genetics*; D012261:Ribonucleoproteins; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D013489:Suppression, Genetic
chemical_list: D001059:Apoproteins; C071249:CBP2 protein, S cerevisiae; D003573:Cytochrome b Group; D005656:Fungal Proteins; D012261:Ribonucleoproteins; D029701:Saccharomyces cerevisiae Proteins; D045303:Cytochromes b