A Novel Form of Transcriptional Silencing by Sum1-1 Requires Hst1 and the Origin Recognition Complex

Abstract

In the yeast Saccharomyces cerevisiae, a and α mating-type information is stored in transcriptionally silenced cassettes called HML and HMR. Silencing of these loci, maintained by the formation of a specialized type of heterochromatin, requires trans-acting proteins andcis-acting elements. Proteins required for silencing include the Sir2 NAD⁺-dependent deacetylase, Sir3, and Sir4. Factors that bind to the cis elements atHMR and HML and that are important for silencing include the origin recognition complex (ORC). Mutations of any of these Sir proteins or combinations of cis elements result in loss of silencing. SUM1-1 was previously identified as a dominant mutation that restores silencing toHMR in the absence of either the Sir proteins or some of the cis elements. We have investigated the novel mechanism whereby Sum1-1 causes Sir-independent silencing at HMR and present the following findings: Sum1-1 requires the Sir2 homolog, Hst1, for silencing and most probably requires the NAD⁺-dependent deacetylase activity of this protein. Sum1-1 interacts strongly with ORC, and this strong interaction is dependent on HMR DNA. Furthermore, ORC is required for Sum1-1-mediated silencing atHMR. These observations lead to a model for Sum1-1 silencing of HMR in which Sum1-1 is recruited toHMR by binding to ORC. Sum1-1, in turn, recruits Hst1. Hst1 then deacetylates histones or other chromatin-associated proteins to cause chromatin condensation and transcriptional silencing.

ACKNOWLEDGMENTS

We thank J. Boeke, L. Pillus, D. Shore, and A. Vershon for plasmids and strains and B. Stillman for the anti-Orc3 antibody. We also thank A. Vershon for communicating results prior to publication.

This work was supported by National Institutes of Health grants GM28220 and GM55641 to R.S. and a Beckman Foundation Scholarship to J.C.