Dia2's F-box and LRR domains are indispensable for transcriptional silencing.

<p>(A) A schematic drawing of the domain structure of Dia2. Dia2 contains a tetratricopeptide repeat (TPR) region, nuclear localization signal (NLS), F-box and leucine rich repeat (LRR) domain. (B) The Dia2 F-box and LRR domains are important for silencing at the <i>HMR</i> locus. The <i>dia2</i>Δ mutant cells containing the <i>HMR::GFP</i> reporter were transformed with empty vector or a plasmid expressing WT or different <i>dia2</i> mutants, and the percentage of cells expressing GFP was determined as in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002846#pgen-1002846-g001" target="_blank">Figure 1B</a>. Note that the percentage of cells expressing GFP in <i>dia2</i>Δ cells was higher when using selective media compared to cells growing in YPD. (C) The Dia2 F-box and LRR domains are important for telomere silencing. The <i>dia2</i>Δ mutants containing the <i>VIIL-URA3</i> reporter were transformed with empty vector or a plasmid expressing the indicated <i>dia2</i> mutant and spotted onto selective media (-HIS) and media containing FOA (-HIS, FOA) to assay silencing. (D–E) The Dia2 LRR domain is important for <i>HMR</i> silencing (D) and telomere silencing (E). Assays were performed as described in B and C.</p>