Coexpression of torsinA and printor does not uncover a phenotype stemming from torsinA expression.

<p>(A) The UPR of the indicated yeast strains was monitored by flow cytometry to detect expression from the UPRE-GFP construct upon stress with CPY* or 1.5 mM DTT. Coexpression of printor does not allow WT torsinA to reduce UPR-related stress. A 1-tailed Student's t-test was used to compare relative fluorescence of torsinA strains to the vector control. * = p<0.01. N = 6 independent trials per each sample. (B) The impact of torsinA with and without printor on trafficking of invertase. Cells were spotted on plates containing the pH sensitive dye, BCP. Simultaneous expression of torsinA and printor does not impact the rate of secretion.</p

TorsinA cannot rescue α-synuclein-induced toxicity in yeast.

<p>Assay showing the ability of yeast to grow in the presence or absence of α-synuclein (α-syn) with and without torsinA. Each row is a 5-fold dilution of the previous row. TorsinA cannot rescue α-syn-induced toxicity.</p

TorsinA can be expressed in the endoplasmic reticulum of yeast.

<p>(A) Diagram of constructs created for this work (upper panel) and summary of the different forms of torsinA used in these experiments (lower panel). TorsinA was localized to the endoplasmic reticulum (ER) using the signal sequence of the endogenous yeast protein KAR2 and an HDEL sequence. (B) Microscopy of yeast strains used. TorsinA was localized to the contiguous lumen of the nuclear envelope and ER. Some signal was also seen in the vacuole (arrow head), suggesting a portion of the protein was degraded. A representative frame is shown for each strain. Scale bar  =  2 µM. (C) Growth of torsinA-expressing yeast on plates. Each row is a 5-fold dilution of the previous row. Expression of wild type (WT) or mutant torsinA did not impact the growth rate. Uninduced plates included 1 mM methionine and induced plates lacked methionine.</p

TorsinA does not impact the unfolded protein response or trafficking in yeast.

<p>(A) A construct containing GFP driven by a unfolded protein response (UPR) sensitive promoter (UPRE-GFP) was used to monitor levels of the unfolded protein response (UPR) upon stress with 1.5 mM dithiothreitol (DTT) or mutant carboxypeptidase Y (CPY*). ERO1 served as a positive control. TorsinA is not able to reduce UPR levels caused by either stressor. Statistical analysis was conducted in comparison to the vector control strain with a 1-tailed Student's t-test. * = p<0.05, # = p<0.005, & = p<0.001. N = 6 independent trials per sample. (B)Growth of ero1-1 in the presence and absence of torsinA at 37°C. Each row is a 5-fold dilution of the previous row. TorsinA is not able to rescue the growth defect by the <i>ero1-1</i> mutation. Uninduced plates included 1 mM methionine and induced plates lacked methionine. (C) Trafficking of invertase, as monitored by halos produced by growth of torsinA-expressing strains on plates containing bromocresol purple (BCP). TorsinA does not impact the rate of secretion.</p