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Fig. 7. The effects of over-expression of the vps1 C-terminal region. (A) The structures of Vps1p and its deletion constructs used in the experiments below. Numbers on the top indicate the amino acid positions. (B) Over-expressions of the C-terminal region of Vps1p caused cell death at 37°C. Wild-type cells containing various deletion constructs of VPS1 (pGAL1-VPS1C1C4, pGAL1-VPS1-N1), as well as the full length VPS1 (pGAL1-VPS1) were grown on glucose containing medium and replicated to galactose plates and incubated at 37°C for one day. (C) The localization of Sla1p was affected by the over-expression of the C-terminal regions of Vps1p. The endogenous copy of SLA1 in wild-type cells was tagged with YFP tag at its C-terminus and the resulting integrant was transformed with various deletion constructs of VPS1 indicated in A. The resulting transformants were cultured in raffinose-containing media to the mid-log phase before galactose was added and incubated for another 3 hours at 30°C. The localization of the various domains of Vps1p along with that of Sla1p was examined under Leica microscopy. Note the partial co-localization of Sla1p with Vps1p and its domains in 1, 4 and 6 (bar, 5 µm). (D) The strains as in (B) were analyzed for their sensitivities to LAT-A. The cells were plated on the galactose-containing medium at 30°C and the LAT-A with the indicated concentrations was applied (left panel). The results were summarized graphically in the right panel.

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