Mutations in a semiconserved region of the Tetrahymena intron

The A-rich bulge in paired region P5a of the Tetrahymena intron is a structural feature that is conserved in the sub-group Ib self-splicing introns. We have constructed a series of substitution and deletion mutations in this region of the intron. Kinetic analysis has shown that some of the mutants have a reduced maximal extent of splicing, while others have a reduced Vmax. These mutations could be reactivated to a great extent by spermidine and high Mg2+ concentrations. These data are consistent with the hypothesis that the A-rich bulge of P5a has a role in stabilizing the higher-level structure of the ribozyme.