Stimulation of Mitotic Recombination Events by High Levels of RNA Polymerase II Transcription in Yeast

Abstract

The impact of high levels of RNA polymerase II transcription on mitotic recombination was examined using lys2 recombination substrates positioned on nonhomologous chromosomes. Substrates were used that could produce Lys⁺ recombinants by either a simple (noncrossover) gene conversion event or a crossover-associated recombination event, by only a simple gene conversion event, or by only a crossover event. Transcription of the lys2 substrates was regulated by the highly inducible GAL1-10 promoter or the low-level LYS2 promoter, with GAL1-10 promoter activity being controlled by the presence or absence of the Gal80p negative regulatory protein. Transcription was found to stimulate recombination in all assays used, but the level of stimulation varied depending on whether only one or both substrates were highly transcribed. In addition, there was an asymmetry in the types of recombination events observed when one substrate versus the other was highly transcribed. Finally, the lys2 substrates were positioned as direct repeats on the same chromosome and were found to exhibit a different recombinational response to high levels of transcription from that exhibited by the repeats on nonhomologous chromosomes. The relevance of these results to the mechanisms of transcription-associated recombination are discussed.

ACKNOWLEDGMENTS

We acknowledge the contributions of Merrilyn Michelitch, Tamara Murphy, Anna Speke, and Miyono Hendrix in constructing some of the substrates and strains used in this analysis. We thank Jennifer Freedman and Takura Nakagawa for helpful comments on the manuscript and members of the laboratory for fruitful discussions.

This work was supported by National Institutes of Health grant GM38464 to S.J.-R. D.S. and A.D. were partially supported by the Emory University Graduate Division of Biological and Biomedical Sciences.