||Two-color cell array screen reveals interdependent roles for histone chaperones and a chromatin boundary regulator in histone gene repression.
||Jeffrey Fillingham, Pinay Kainth, Jean-Philippe Lambert, Harm van Bakel, Kyle Tsui, Lourdes Peña-Castillo, Corey Nislow, Daniel Figeys, Timothy R Hughes, Jack Greenblatt, Brenda J Andrews
||We describe a fluorescent reporter system that exploits the functional genomic tools available in budding yeast to systematically assess consequences of genetic perturbations on gene expression. We used our Reporter-Synthetic Genetic Array (R-SGA) method to screen for regulators of core histone gene expression. We discovered that the histone chaperone Rtt106 functions in a pathway with two other chaperones, Asf1 and the HIR complex, to create a repressive chromatin structure at core histone promoters. We found that activation of histone (HTA1) gene expression involves both relief of Rtt106-mediated repression by the activity of the histone acetyltransferase Rtt109 and restriction of Rtt106 to the promoter region by the bromodomain-containing protein Yta7. We propose that the maintenance of Asf1/HIR/Rtt106-mediated repressive chromatin domains is the primary mechanism of cell-cycle regulation of histone promoters. Our data suggest that this pathway may represent a chromatin regulatory mechanism that is broadly used across the genome.
||Mol. Cell 2009; 35:340-51
||The promoter sequence of HTA is fused to GFP along with a control reporter construct with the constitutively expressed RPL39 promoter driving tdTomato (RFP) expression.
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