Abstract
Ume1p is a member of a conserved protein family including RbAp48 that associates with histone deacetylases. Consistent with this finding, Ume1p is required for the full repression of a subset of meiotic genes during vegetative growth in budding yeast. In addition to mitotic cell division, this report describes a new role for Ume1p in meiotic gene repression in precommitment sporulating cultures returning to vegetative growth. However, Ume1p is not required to re-establish repression as part of the meiotic transient transcription program. Mutational analysis revealed that two conserved domains (NEE box and a WD repeat motif) are required for Ume1p-dependent repression. Co-immunoprecipitation studies revealed that both the NEE box and the WD repeat motif are essential for normal Rpd3p binding. Finally, Ume1p-Rpd3p association is dependent on the global co-repressor Sin3p. Moreover, this activity was localized to one of the four paired amphipathic-helix domains of Sin3p shown previously to be required for transcriptional repression. These findings support a model that Ume1p binding to Rpd3p is required for its repression activity. In addition, these results suggest that Rpd3-Ume1p-Sin3p comprises an interdependent complex required for mediating transcriptional repression.
Highlights
The second repression system operates through the URS1 promoter element found in several early meiotic genes [11,12,13,14]
In addition to mitotic cell division, this report describes a new role for Ume1p in meiotic gene repression in precommitment sporulating cultures returning to vegetative growth
We demonstrate that Ume1p is required for early meiotic gene repression in vegetative cultures regardless of the carbon source and in precommitment meiotic cells returning to mitotic cell division
Summary
All growth and sporulation procedures have been described [25]. Meiotic progression was followed by staining samples with the DNAspecific stain 4Ј,6-diamidino-2-phenylindole as described [26]. The completion of meiosis I and meiosis II was determined by the appearance of binucleated and tetranucleated cells, respectively. Plasmids and S1 Protection Probes—The plasmid containing the spo13-lacZ reporter gene (p(spo13)28) has been described elsewhere.
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