Figure 5.

Promiscuous Mst2 Activity in the Absence of Pdp3 Attenuates Heterochromatin Silencing

(A) Mst2 DamID maps for the centromere of chromosome 1 in WT, pdp3Δ, and set2Δ cells. The signal of DamMst2 (normalized to Dam only) was averaged over 500 probes and is shown in log₂ scale. The x axis shows position on chromosomes.

(B and D) RNA expression in WT, pdp3Δ, and mst2Δ cells at centromere 1 (B) and telomere 1L (D) assessed by RNA-seq. Relative read counts normalized to total read number (axis scale in log₂) are shown.

(C) Relative RNA expression levels of ura4⁺ (qRT-PCR analysis) at the innermost repeat (imr) in indicated mutants. Shown are the transcript levels relative to WT after normalization to act1⁺. Data are represented as mean ± SEM from four independent biological experiments.

(E) Scheme of experimental setup for tethering Mst2 at heterochromatin: Four LexA-binding sites were inserted downstream of the ade6⁺ reporter at the outermost repeat (otr) of chromosome 1 (top). The ade6-M210 allele at the euchromatic endogenous locus of ade6 (bottom) was used as a reference.

(F–H) Expression levels of heterochromatic ade6⁺ reporter relative to euchromatic ade6-M210 allele in WT (F), pdp3Δ (G), or set2Δ (H) cells. The p values were calculated using the two-sided, two-sample Student’s t test. Error bars indicate SD (n ≥ 3 independent biological replicates).

See also Figure S2.