Removal of Frameshift Intermediates by Mismatch Repair Proteins in Saccharomyces cerevisiae

Abstract

Frameshift mutations occur when the coding region of a gene is altered by addition or deletion of a number of base pairs that is not a multiple of three. The occurrence of a deletion versus an insertion type of frameshift depends on the nature of the transient intermediate structure formed during DNA synthesis. Extrahelical bases on the template strand give rise to deletions, whereas extrahelical bases on the strand being synthesized produce insertions. We previously used reversion of a +1 frameshift mutation to analyze the role of the mismatch repair (MMR) machinery in correcting −1 frameshift intermediates within a defined region of the yeast LYS2gene. In this study, we have used reversion of a −1 frameshift mutation within the same region of LYS2 to analyze the role of the MMR machinery in the correction of frameshift intermediates that give rise to insertion events. We found that insertion and deletion events occur at similar rates but that the reversion spectra are very different in both the wild-type and MMR-defective backgrounds. In addition, analysis of the +1 spectra revealed novel roles for Msh3p and Msh6p in removing specific types of frameshift intermediates.

ACKNOWLEDGMENTS

We gratefully acknowledge the contributions of C. Greene and N. Morey in constructing and generously donating strain SJR922 for use in this study. We thank K. Hill, G. F. Crouse, and members of the S.J.-R. lab for critically reading the manuscript.

This work was funded by a grant from the National Science Foundation.