Article abstract
Nature Genetics 41, 800 - 806 (2009)
Published online: 21 June 2009 | doi:10.1038/ng.402
The establishment of gene silencing at single-cell resolution
Erin A Osborne1, Sandrine Dudoit2,3 & Jasper Rine3,4
Abstract
The establishment of silencing in Saccharomyces cerevisiae is similar to heterochromatin formation in multicellular eukaryotes. Previous batch culture studies determined that the de novo establishment of silencing initiates during S phase and continues for up to five cell divisions for completion. To track silencing phenotypically, we developed an assay that introduces Sir3 protein into individual sir3
mutant cells synchronously and then detects the onset of silencing with single-cell resolution. Silencing was completed within the first one to two cell divisions in most cells queried. Moreover, we uncovered unexpected complexity in the contributions of a histone acetyltransferase (Sas2), two histone methytransferases (Dot1 and Set1) and one histone demethylase (Jhd2) to the dynamics of silencing. Our findings showed that removal of methyl modifications at H3K4 and H3K79 were important steps in silent chromatin formation and that Jhd2 and Set1 had competing roles in the process.
- Department of Plant and Microbial Biology, University of California, Berkeley, Berkeley, California, USA.
- Division of Biostatistics at the School of Public Health, University of California, Berkeley, Berkeley, California, USA.
- California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, California, USA.
- Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA.
Correspondence to: Jasper Rine3,4 e-mail: jrine@berkeley.edu
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