Circular Minichromosomes Become Highly Recombinogenic in Topoisomerase-deficient Yeast Cells

Abstract

In topoisomerase-deficient yeast cells, we have found that circular minichromosomes are present as broad distributions of multimeric forms, which consist of tandemly repeated copies of their monomeric sequences. This phenomenon selectively occurs in Deltatop1 cells, and is highly magnified in double mutant Deltatop1 top2-4 cells. No multimers are observed in single mutant top2-4 or Deltatop3 cells, or in Deltatop1 cells that express a plasmid-borne TOP1 gene. Interconversion among multimeric forms takes place rapidly in double mutant Deltatop1 top2-4 cells, and the multimeric distributions are readily reverted to the monomeric form when a plasmid-borne TOP1 gene is expressed from an inducible promoter. These observations are a new example of the interplay between DNA topology and genome stability, and suggest that the cell capacity to modulate DNA supercoiling is limited when DNA is organized in small topological domains. Yeast minichromosome multimerization provides an appropriate system in which to study mechanistic aspects of DNA recombination.