Substitution as a Mechanism for Genetic Robustness: The Duplicated Deacetylases Hst1p and Sir2p in Saccharomyces cerevisiae

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Abstract
How duplicate genes provide genetic robustness remains an unresolved question. We have examined the duplicated histone deacetylases Sir2p and Hst1p in Saccharomyces cerevisiae and find that these paralogs with non-overlapping functions can provide genetic robustness against null mutations through a substitution mechanism. Hst1p is an NAD+-dependent histone deacetylase that acts with Sum1p to repress a subset of midsporulation genes. However, hst1Δ mutants show much weaker derepression of target loci than sum1Δ mutants. We show that this modest derepression of target loci in hst1Δ strains occurs in part because Sir2p substitutes for Hst1p. Sir2p contributes to repression of the midsporulation genes only in the absence of Hst1p and is recruited to target promoters by a physical interaction with the Sum1 complex. Furthermore, when Sir2p associates with the Sum1 complex, the complex continues to repress in a promoter-specific manner and does not spread. Our results imply that after the duplication, SIR2 and HST1 subfunctionalized. The single SIR2/HST1 gene from Kluyveromyces lactis, a closely related species that diverged prior to the duplication, can suppress an hst1Δ mutation in S. cerevisiae as well as interact with Sir4p in S. cerevisiae. In addition, the existence of two distinct protein interaction domains for the Sir and Sum1 complexes was revealed through the analysis of a chimeric Sir2–Hst1 molecule. Therefore, the ability of Sir2p to substitute for Hst1p probably results from a retained but reduced affinity for the Sum1 complex that is a consequence of subfunctionalization via the duplication, degeneration, and complementation mechanism. These results suggest that the evolutionary path of duplicate gene preservation may be an important indicator for the ability of duplicated genes to contribute to genetic robustness. Gene duplication is an important force in evolution, as it provides a source of new genetic material. However, the mechanisms by which duplicated genes are retained and diverge are understudied at the experimental level. We have examined a pair of duplicated histone deacetylases Hst1p and Sir2p from baker's yeast, which are important for distinct types of gene repression. In this study, we show that before the duplication the ancestral histone deacetylase had both Hst1p- and Sir2p-like functions, and after the duplication Sir2p and Hst1p subfunctionalized, giving rise to two distinct proteins with normally nonoverlapping functions. Despite having partitioned the ancestral functions after the duplication, Sir2p can substitute for Hst1p in its absence by interacting with the normal partner of Hst1p. This study suggests that the evolutionary path of duplicate gene preservation may be an important indicator for the ability of duplicated genes to substitute for one another and hence protect the organism against deleterious mutations.