The URA3-tel, GF2 and GF3 constructs (shown on top) were integrated in the strains shown on the left and the level of URA3 silencing was calculated as %FOA^R cells. A) Levels of URA3 silencing (%FOA^R). The 0-10% range is spread out to properly show differences at very low levels of silencing. B) Ratios of %FOA^R in the mutant strains versus the %FOA^R in the wild type (BY4742) strain. Data is from Table S1. The arrows underneath the exponential graph indicate increase or decrease of silencing.

The URA3-tel, GF2, GF3, GF6, GF9 and GF46 constructs (shown on top) were integrated in the strains shown on the left and the level of URA3 silencing was calculated as %FOA^R cells. A) Levels of URA3 silencing (%FOA^R). The 0-10% range is spread out to properly show differences at very low levels of silencing. B) Ratios of %FOA^R in the mutant strains versus the %FOA^R in the wild type strain. Wild type depicts BY474 for Δrif1 and W303 (not shown) for cdc6-1. The arrows underneath the exponential graph indicate increase or decrease of silencing.

The URA3-tel, GF2, GF6 and GF6ΔACS constructs (shown on top) were integrated in the strains shown along the vertical axis and the level of URA3 silencing was calculated as %FOA^R cells. A) Levels of URA3 silencing (%FOA^R). The 0-10% range in the upper graph is spread out to properly show differences at very low levels of silencing. The levels of silencing of STAR-containing (grey bars) and STAR-less (black bars) constructs are shown side by side. B) Ratios of %FOA^R in STAR-less versus STAR-containing constructs. The ratios %FOA^R_URA3-tel/%FOA^R_GF2 and %FOA^R_GF6ΔSTAR/%FOA^R_GF6 were calculated and plotted. The arrows underneath the exponential graph indicate increase or decrease of silencing.

A) Telomeric reporters used in this study. Maps (not to scale) of the used constructs are shown. The positions of core X element from the IIR telomere and the Y' element from the XII-L telomere (black rectangles), the STARs from the same telomeres (grey rectangles), URA3, ADH4 and the telomeric TG_1-3 repeats (black triangles) are as indicated. The position of the destroyed ACS (ARS Consensus Sequence) is depicted by an open diamond. The 5′→3′ direction of URA3 transcription is indicated in the URA3-tel construct (top) and is the same for all constructs shown. The insertions between URA3 and the telomeric repeat add 145-900 base pairs as compared to URA3-tel. B) Percentage of FOA^R cells in different strains and constructs. The reporter constructs shown along the vertical axis were integrated in the strains shown on the left. Percentage of FOA^R cells was measured in at least three independent experiments. Average %FOA^R ± std. dev. were calculated and plotted. Data is from Table S1. C) URA3-tel recapitulates silencing effects in mutant strains. The URA3-tel construct was integrated in the strains shown along the vertical axis. The ratios of %FOA^R in the mutant strains versus the %FOA^R in the isogenic wild type strain were calculated and plotted. The effects of Δsas2, Δsas3, Δyng1, Δrtt109, Δhat1, Δgcn5 and Δrif1 were assessed using BY4742 as the wild type strain (Table S1). The effects of orc2-1, orc5-1, mcm5-461, cdc6-1, cdc45-1 and cdc7-1(sas1) were assessed using W303 as the wild type strain (data not shown). There is little difference in the levels of telomeric silencing between BY4742 and W303. The arrows underneath the exponential graph indicate increase or decrease of silencing.

The GF6, GF9, GF44, GF46 and GF61 constructs (shown on top) were integrated in the strains shown on the left and the level of URA3 silencing was calculated as %FOA^R cells. A) Levels of URA3 silencing (%FOA^R). B) Ratios of %FOA^R in the mutant strains versus the %FOA^R in the wild type (BY4742) strain. Data is from Table S1. The arrows underneath the exponential graph indicate increase or decrease of silencing.

GF6, GF6ΔACS, GF44 and GF6ΔACS constructs (shown on the left) were integrated in the strains shown along the vertical axis and the level of URA3 silencing was calculated as %FOA^R cells. A) Levels of URA3 silencing (%FOA^R). The levels of silencing of ACS-containing (black bars) and ACS-less (grey bars) constructs are shown side by side. B) Ratios of %FOA^R in ACS-less versus ACS-containing constructs. The ratios %FOA^R_GF6ΔACS/%FOA^R_GF6 and %FOA^R_GF44ΔACS/%FOA^R_GF44 were calculated and plotted. The arrows underneath the exponential graph indicate increase or decrease of silencing.