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Science 26 January 2001:
Vol. 291. no. 5504, pp. 650 - 653
DOI: 10.1126/science.291.5504.650
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Reports

Establishment of Transcriptional Silencing in the Absence of DNA Replication

Yao-Cheng Li, Tzu-Hao Cheng,* Marc R. Gartenbergdagger ddagger

Transcriptional repression of the silent mating-type loci in Saccharomyces cerevisiae requires a cell cycle-dependent establishment step that is commonly assumed to involve DNA replication. Using site-specific recombination, we created a nonreplicating DNA ring in vivo to test directly the role of replication in establishment of silencing. Sir1 was tethered to the ring following excision from the chromosome to activate a dormant silencer. We show here that silencing can be established in DNA that does not replicate. The silenced ring adopted structural features characteristic of bona fide silent chromatin, including an altered level of DNA supercoiling and reduced histone acetylation. In addition, the process required silencing factors Sir2, Sir3, and Sir4 and progression between early S and M phases of the cell cycle. The results indicate that passage of a replication fork is not the cell-cycle event required for establishment of silencing in yeast.

Department of Pharmacology, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, Piscataway, NJ 08854, USA.
*   Present address: Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.

dagger    Member, The Cancer Institute of New Jersey, New Brunswick, NJ 08901, USA.

ddagger    To whom correspondence should be addressed. E-mail: gartenbe{at}umdnj.edu

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