Supplemental Material for Edskes, et al., 2021

The Supplementary Information includes extended SILAC methods, strains of <i>S. cerevisiae </i>(Table S1), primers used (Table S2), proteins increased 1.5-fold or more in abundance in <i>pre9</i>Δ (Table S3), Proteins decreased 1.5-fold or more in abundance in <i>pre9</i>Δ (Table S4), a diagram of the proteasome α ring (Fig. S1), evidence that proteasome – related knockouts do not affect [PSI+] propagation (Fig. S2), evidence that [URE3-1] is less stable in 779-6A <i>pre</i>9Δ than in BY241 <i>pre9</i>Δ (Fig. S3), data showing that levels of Sup35p are not affected by proteasome – related mutations in strain BY241 (Fig. S4), data showing Btn2p levels in proteasome mutants are elevated by the presence of [PSI+] (Fig. S5 and S6), measuring the absolute levels of Btn2p and Cur1p in strain 779-6A by comparison with purified recombinant Btn2-His6 and Cur1-His6 (Fig. S7), the multiple bands of Cur1p (Fig. S8), the elevation of Btn2p in <i>tof2</i> mutants (Fig. S9) and the absence of a ubiquitin ladder of Btn2p in a <i>pre9</i>Δ strain (Fig. S10). Table S5 compares protein turnover rates with their levels by SILAC in <i>pre9</i>Δ strains.

本补充信息包含经扩展的稳定同位素标记氨基酸细胞培养（Stable Isotope Labeling by Amino acids in Cell culture，SILAC）实验方法、酿酒酵母（S. cerevisiae）菌株（表S1）、所用引物（表S2）、pre9Δ菌株中丰度上调1.5倍及以上的蛋白质（表S3）、pre9Δ菌株中丰度下调1.5倍及以上的蛋白质（表S4）；此外还包含蛋白酶体α环结构示意图（图S1）、蛋白酶体相关基因敲除不影响[PSI+]传播的验证结果（图S2）、[URE3-1]在779-6A pre9Δ菌株中的稳定性低于BY241 pre9Δ菌株的实验证据（图S3）、BY241菌株中Sup35p蛋白水平不受蛋白酶体相关突变影响的实验数据（图S4）、蛋白酶体突变体中Btn2p蛋白水平因[PSI+]存在而上调的实验数据（图S5与S6）；通过与纯化的重组Btn2-His6和Cur1-His6蛋白进行定量对比，测定779-6A菌株中Btn2p与Cur1p的绝对表达量（图S7）、Cur1p的多条带现象（图S8）、tof2突变体中Btn2p的表达上调（图S9）以及pre9Δ菌株中未检测到Btn2p的泛素梯带（图S10）。表S5对比了pre9Δ菌株内蛋白质的周转速率与其通过SILAC测得的丰度水平。