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The DNA Repair Protein yKu80 Regulates the Function of Recombination Enhancer during Yeast Mating Type Switching

Huck Institutes of the Life Sciences, Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802,¹ Stowers Institute for Medical Research, Kansas City, Missouri 64110²

Received 14 February 2005/ Returned for modification 24 March 2005/ Accepted 11 July 2005

Recombination enhancer (RE) is essential for regulating donor preference during yeast mating type switching. In this study, by using minichromosome affinity purification (MAP) and mass spectrometry, we found that yeast Ku80p is associated with RE in MATa cells. Chromatin immunoprecipitation assays confirmed its occupancy in vivo. Deletion of YKU80 results in altered chromatin structure in the RE region and more importantly causes a dramatic decrease of HML usage in MATa cells. We also detect directional movement of yKu80p from the RE towards HML during switching. These results indicate a novel function of yeast Ku80p in regulating mating type switching.

This manuscript is dedicated to the memory of Robert T. Simpson, a trusted colleague and thoughtful mentor.

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