Regulation of Nuclear Positioning and Dynamics of the Silent Mating Type Loci by the Yeast Ku70/Ku80 Complex

Abstract

ABSTRACT We have examined the hypothesis that the highly selective recombination of an active mating type locus ( MAT ) with either HML α or HMR a is facilitated by the spatial positioning of relevant sequences within the budding yeast ( Saccharomyces cerevisiae ) nucleus. However, both position relative to the nuclear envelope (NE) and the subnuclear mobility of fluorescently tagged MAT , HML , or HMR loci are largely identical in haploid a and α cells. Irrespective of mating type, the expressed MAT locus is highly mobile within the nuclear lumen, while silent loci move less and are found preferentially near the NE. The perinuclear positions of HMR and HML are strongly compromised in strains lacking the Silent information regulator, Sir4. However, HML α, unlike HMR a and most telomeres, shows increased NE association in a strain lacking yeast Ku70 (yKu70). Intriguingly, we find that the yKu complex is associated with HML and HMR sequences in a mating-type-specific manner. Its abundance decreases at the HML α donor locus and increases transiently at MAT a following DSB induction. Our data suggest that mating-type-specific binding of yKu to HML α creates a local chromatin structure competent for recombination, which cooperates with the recombination enhancer to direct donor choice for gene conversion of the MAT a locus.