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Fig. 6. Overexpression of Nvj1p, but not a mutant form lacking Osh1p-binding activity, confers a tryptophan transport-related growth defect. (A) Cells overexpressing Nvj1p, but not a mutant form lacking the Osh1p-binding domain, grow poorly on media containing limiting concentrations of tryptophan at low temperature. Cells harboring P_CUP1-NVJ1, P_CUP1-NVJ1(130-176aa)-myc, or empty vector were induced and replica-plated onto media containing limiting (15 µg/ml) or excess (40 µg/ml) tryptophan as described in Materials and Methods. (B) The cold-sensitive growth phenotype of Nvj1p-overexpressing cells is suppressed by low- and high-affinity tryptophan permeases. Cells harboring P_CUP1-NVJ1 and either pTAT1, pTAT2, or empty vector were induced and replica-plated onto media containing limiting (15 µg/ml) or excess (40 µg/ml) tryptophan as above. (C) NV junctions are not necessary to confer tryptophan transport defects in Nvj1p-overexpressing cells. Growth curves of vac8- cells expressing P_CUP1-NVJ1 (open squares), P_CUP1-NVJ1(130-176aa)-myc (black triangles), or empty vector (gray circles) were determined in liquid media containing limiting (15 µg/ml) or excess (40 µg/ml) tryptophan at 25°C using a Bioscreen C Analyzer (see Materials and Methods).

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