A Yeast Model of FUS/TLS-Dependent Cytotoxicity
Figure 3
Toxicity and localization of individual domains of FUS/TLS.
(A) A serial deletion of the full length FUS/TLS gene from c-terminus, labeled as 1–4, and from N-terminus, labeled as 6–8, was carried out. (B) The truncated genes (with GFP tag at N-terminus) were then placed under the control of GAL1 promoter on the yeast expression vector pYES2CT. Yeast with above constructs was serially diluted and spotted onto plate containing either glucose (expression “off”) or galactose (expression “on”). Pictures of the plates were taken after 2 d growth at 30°C. (C) Cells containing the above constructs were grown in the Ura-Raffinose medium to mid-log phase. Expression of the proteins was induced by 2% galactose for 6 h. Localization and aggregation of the proteins was visualized by fluorescence microscopy. (D) Constructs 3–5 as shown in (A) were also cloned into yeast expression vector pDEST52 without the GFP tag. Yeast containing the constructs was serially diluted and spotted onto glucose (expression “off”) or galactose plate (expression “on”). Picture of the plates was taken after 2 d growth at 30°C.