![]() ![]() The contributions of each mutation to the genetic interaction and the strong position specificity of suppression, combined with previous findings, support a model in which the 5' /GUA and the GAG of U6 function in binding the 3' YAG/ during the second catalytic step. RNA analysis confirmed that the severe splicing defect observed in A+3 and Y-3 double mutants can be rescued to near wild-type levels by the mutations in U6 G50. This suppression is significantly enhanced upon the inclusion of a specific mutation Y-3 in the 3' YAG/. ![]() There was no striking position-dependence of individual local consensus sequences when viewed over the entire population this contrasted starkly with the SD motifs, which were much more position-dependent. Two mutations in U6 G50 of the ACAGAG can weakly suppress two mutations in A+3 of the 5' /GUA. Instead, shorter, significant (FDR<0.01) motifs that were analyzed independently of the 16S rRNA comprise many local consensus sequences. Here we report several striking genetic interactions between A+3 of the 5' /GUA with Y-3 of the 3' YAG/ and G50 of the highly conserved ACAGAG motif in U6 snRNA. Although previous observations have suggested that the G of the YAG/ interacts with the first nucleotide of the /GUA consensus sequence at the 5' end of the intron, additional interactions have not been identified. Little is known about the interactions formed by these three nucleotides in the spliceosome. The YAG/ consensus sequence at the 3' end of introns (the slash indicates the location of the 3' splice site) is essential for catalysis of the second step of pre-mRNA splicing. When similarity was quantified by a consensus value, both extremely low and extremely high values were notably absent from the wild-type sequences of the. ![]()
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