In transcriptional silencing, SIR2,3,4 are required in stoichiometric amounts to silence specific chromosomal regions. In yeast, SIR proteins bind sites on nucleosome tails and form a multimeric compound of SIR2,3,4 that condenses chromatin and is thought to physically occlude promoters in the silenced interval, preventing their interaction with transcription machinery. The establishment of SIR-repressed heterochromatin domains is a complicated process that involves different subsets of proteins and regulatory proteins depending on the locus in the genome. At the silent mating type loci and at yeast telomeres, the transcription factors Abf1 (ARS binding factor) and Rap1 (repressor-activator protein) associate with specific nucleotide sequences in the silencers that flank heterochromatic regions. Rap1 contains a Sir3-binding domain that recruits SIR3 to the silencers. Once at the silencers, Sir3 recruits Sir4-Sir2 dimers to the chromatin nucleation site. Sir2 then deacetylates histone H3 and H4 tails, and free Sir3 binds the now-deacetylated lysine residues H4K16,79, and recruits additional Sir4-Sir2 dimers to promote the further spreading of the heterochromatin domain.
Once it has spread to cover a genomic locus, the SIR2,3,4 effectively prevents transcription from the region it occupies, in a process that is thought to depend on the physical occlusion of DNA by SIR proteins. Recently, it has been shown that certain promoters are capable of directing transcription inside regions that are otherwise silenced by SIR proteins. Specifically, if an inducible promoter is induced inside a silent chromatin domain, it can achieve ~200x increase in expression levels with little detectable change in covalent histone modifications.