Activation of a cryptic TACTAAC box in the Saccharomyces cerevisiae actin intron.
We constructed a translational fusion between the Saccharomyces cerevisiae actin gene and the Escherichia coli beta-galactosidase structural gene such that expression of beta-galactosidase activity required accurate splicing of the actin intron. Using this chimeric gene, we generated a series of internal deletions which removed the TACTAAC box or, in addition, TACTAAC-like sequences within the intron. Analysis of the fusion transcripts produced in these deletions allowed us to conclude that the TACTAAC-like sequence TACTAAG can substitute, albeit inefficiently, for the authentic TACTAAC box in the splicing process. These results indicate that the yeast splicing machinery can utilize a cryptic TACTAAC box, but there are requirements for primary sequence and proper position.
D000199:Actins / Q000235:genetics*; D001483:Base Sequence; D002872:Chromosome Deletion; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D005798:Genes, Bacterial; D005800:Genes, Fungal; D010957:Plasmids; D014176:Protein Biosynthesis; D012326:RNA Splicing; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D000199:Actins; D004262:DNA Restriction Enzymes; D001616:beta-Galactosidase