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Originally published as Genetics Published Articles Ahead of Print on June 18, 2006.
Genetics, Vol. 173, 1939-1950, August 2006, Copyright © 2006
doi:10.1534/genetics.106.055491
New Alleles of SIR2 Define Cell-Cycle-Specific Silencing Functions
Mirela Matecic*,1,2,
Kristen Martins-Taylor*,1,
Merrit Hickman*,3,
Jason Tanny
,4,
Danesh Moazed and
Scott G. Holmes*,5
* Department of Molecular Biology and Biochemistry, Wesleyan University, Middletown, Connnecticut 06459 and Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115
5 Corresponding author: Department of Molecular Biology and Biochemistry, Hall-Atwater Laboratories, Lawn Ave., Wesleyan University, Middletown, CT 06459-0175.
E-mail: sholmes{at}wesleyan.edu
The establishment of transcriptional silencing in yeast requires cell-cycle progression, but the nature of this requirement is unknown. Sir2 is a protein deacetylase that is required for gene silencing in yeast. We have used temperature-sensitive alleles of the SIR2 gene to assess Sir2's contribution to silencing as a function of the cell cycle. When examined in vivo, these conditional alleles fall into two classes: one class exhibits a loss of silencing when raised to the nonpermissive temperature regardless of cell-cycle position, while the second class exhibits a mitosis-specific silencing defect. Alleles of the first class have a primary defect in protein deacetylase activity, while the alleles of the second class are specifically defective in Sir2–Sir4 interactions at nonpermissive temperatures. Using a SIR2 temperature-sensitive allele, we show that silencing can be established at the HML locus during progression through the G2/M–G1 interval. These results suggest that yeast heterochromatin undergoes structural transitions as a function of the cell cycle and support the existence of a critical assembly step for silent chromatin in mitosis.
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