Copy number and stability of yeast 2 mu-based plasmids carrying a transcription-conditional centromere

Abstract

For certain yeast plasmids, the presence of a centromere segment (CEN) enhances mitotic stability and results in low copy number. Transcription from an inducible promoter adjacent to a CEN segment has been shown to alter centromere function. The rate of loss of a conditional CEN-ARS plasmid was examined and the results suggest that segregation control was immediately and effectively inactivated upon shift to inducing conditions. The effect of a conditional centromere on stability and copy number of hybrid CEN3-2 mu plasmids was also examined. When transcription was repressed, copy number was low. Mitotic stability varied and was correlated with the presence of an intact 2 mu recombination system. When transcription was induced, plasmid copy number increased. However, plasmids became highly unstable. These results indicate that while centromere function is affected by transcription from an adjacent promoter the centromere remains incompatible with the 2 mu maintenance system and may retain partial function.