Characterization of responsiveness across different parameters highlights the factors influencing the C′ AID-GFP efficiency. (A) Fluorescent images of selected proteins were imaged after no (0 min), short (30 min), or long (24 h) periods of induction. Different strains respond to the treatment with different kinetics, with some being depleted below detection after short induction times (Lia1), while others require longer induction times but were below detection levels after 24 h of induction (Mrh1). In cases where the signal is too dim, the outline of the cells is depicted with a dotted line. Scale bar: 5 µm. (B) Dot plot showing the mean GFP fluorescence intensity per cell for each strain before or after a short induction time. The background autofluorescence is marked in dotted lines (A.F., mean and two standard deviations in each direction). The diagonal line indicates the expected location for irresponsive strains. Already in this short induction time, there is a clear response in many strains. (C and D) Schematics depicting the differences in responsiveness for proteins that localize to mitochondria (panel C) and are co- versus posttranslationally translocated (Williams et al., 2014), or to the endoplasmic reticulum (panel D) and have their C′ exposed in the cytosol or hidden in the lumen. Proteins targeted to mitochondria only after their synthesis is complete (post translational) are more likely to be depleted below detection levels upon induction of the AID system. Proteins with the C′ exposed to the cytosol are also more likely to be degraded than their counterparts with hidden C′. This reflects differences in the accessibility of the AID tag to the TIR1 adaptor protein, the ubiquitination machinery and the proteasome, required for its induced depletion. Translocation and topology information were taken from the literature (Jan et al., 2014; Weill et al., 2019). (E) Bar plot of percentage of fluorescent strains versus abundance decile, with decile 1 being the most abundant proteins (Nash et al., 2020). As expected, abundant proteins are more likely to have a fluorescent signal. (F) Bar plot of percentage of responsive strains versus abundance decile (Nash et al., 2020). For the fluorescent strains in each decile, no clear correlation between abundance and responsiveness is detected. Responsiveness remains above 80% for all the deciles.
Figure S2.

Characterization of responsiveness across different parameters highlights the factors influencing the C′ AID-GFP efficiency. (A) Fluorescent images of selected proteins were imaged after no (0 min), short (30 min), or long (24 h) periods of induction. Different strains respond to the treatment with different kinetics, with some being depleted below detection after short induction times (Lia1), while others require longer induction times but were below detection levels after 24 h of induction (Mrh1). In cases where the signal is too dim, the outline of the cells is depicted with a dotted line. Scale bar: 5 µm. (B) Dot plot showing the mean GFP fluorescence intensity per cell for each strain before or after a short induction time. The background autofluorescence is marked in dotted lines (A.F., mean and two standard deviations in each direction). The diagonal line indicates the expected location for irresponsive strains. Already in this short induction time, there is a clear response in many strains. (C and D) Schematics depicting the differences in responsiveness for proteins that localize to mitochondria (panel C) and are co- versus posttranslationally translocated (Williams et al., 2014), or to the endoplasmic reticulum (panel D) and have their C′ exposed in the cytosol or hidden in the lumen. Proteins targeted to mitochondria only after their synthesis is complete (post translational) are more likely to be depleted below detection levels upon induction of the AID system. Proteins with the C′ exposed to the cytosol are also more likely to be degraded than their counterparts with hidden C′. This reflects differences in the accessibility of the AID tag to the TIR1 adaptor protein, the ubiquitination machinery and the proteasome, required for its induced depletion. Translocation and topology information were taken from the literature (Jan et al., 2014; Weill et al., 2019). (E) Bar plot of percentage of fluorescent strains versus abundance decile, with decile 1 being the most abundant proteins (Nash et al., 2020). As expected, abundant proteins are more likely to have a fluorescent signal. (F) Bar plot of percentage of responsive strains versus abundance decile (Nash et al., 2020). For the fluorescent strains in each decile, no clear correlation between abundance and responsiveness is detected. Responsiveness remains above 80% for all the deciles.

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