The Conserved Terminal Guanosine of a Group I Intron can Help Prevent Reopening of the Ligated Exons

Abstract

The highly conserved terminal guanosine of the Tetrahymena group I intron was mutated to an adenosine. The intron still excised itself but more slowly than the wild-type. At very low concentrations of GTP only ligated exons were produced, but at high concentrations of GTP unligated exons accumulated and the 3′ exon acquired a GTP at its 5′ end. A series of experiments suggested that GTP was primarily reopening the ligated exons, rather than directly cleaving the 3′ splice site. 5′ Truncated forms of the wild-type intron can cleave RNA in trans. Cleavage takes place downstream of sequences similar to the last few bases of the 5′ exon. The ligated exons would therefore be a potential substrate. We believe that the intron uses the terminal guanosine to compete with exogenous GTP until the ligated exons have dissociated from their binding site. Other interactions between intron sequences which are known to aid in 3′ splice-site recognition may assist in this process.