Construction of a yeast actin gene intron deletion mutant that is defective in splicing and leads to the accumulation of precursor RNA in transformed yeast cells.

Abstract

The actin gene in yeast Saccharomyces cerevisiae is interrupted by a 309-base-pair intron within the protein-coding region. By using nuclease BAL-31, several intron deletion mutants were constructed to define sequences at the 5' splice junction that are required for RNA splicing. Extensive parts of the intron can be removed without affecting correct splicing. One mutant gene from which the invariant thymidine residue in the second intron position was deleted led to the accumulation of large amounts of unspliced actin mRNA when introduced into yeast cells through a recombinant high-copy-number plasmid. No evidence for the usage of alternative splice sites was obtained.