Limits to transcriptional silencing in Saccharomyces cerevisiae

Mating-type switching in the budding yeast Saccharomyces cerevisiae relies on the Sir protein complex to silence HML and HMR, the two loci containing copies of the alleles of the mating type locus, MAT. Sir-based transcriptional silencing has been considered locus-specific, but the recent discovery of rare and transient escapes from silencing at HML alpha 2 with a sensitive assay called to question if these events extend to the whole locus. Adapting the same assay, we measured that transient silencing failures at HML were more frequent for the alpha 2 gene than alpha 1, similarly to their expression level in unsilenced cells. By coupling a mating assay, at HML we found that one of the two genes at that locus can be transiently expressed while the other gene is maintained silent. Thus, transient silencing loss can be a property of the gene rather than the locus. Cells lacking the SIR1 gene experience epigenetic bistability at HML and HMR. Our previous result led us to ask if HML could allow for two independent epigenetic states within the locus in a sir1 Delta mutant. A simple construct using a double fluorescent reporter at HML alpha 1 and HML alpha 2 ruled out this possibility. Each HML locus displayed a single epigenetic state. We revisited the question of the correlation between the states of two HML loci in diploid cells, and showed they were independent. Finally, we determined the relative strength of gene repression achieved by Sir-based silencing with that achieved by the a1-alpha 2 repressor.

Automated summary

Mating-type switching in Saccharomyces cerevisiae relies on the Sir protein complex for silencing. The recent discovery of rare escapes from silencing raised questions about the extent of this phenomenon. Our study found that transient silencing failures at HML were more frequent for the alpha 2 gene, suggesting a gene-specific rather than locus-wide effect.