FIGURE 6:

The zinc finger of Lnp1 is not required for NPC function. (A) Parental cells exogenously expressing either Lnp1-GFP or lnp1Δznfn-GFP were grown to early log phase at 25°C and visualized by fluorescence microscopy. Scale bar, 5 μm. (B) Expression of lnp1Δznfn results in rescue of lnp1Δ nup133Δ. The lnp1Δ nup133Δ mutants were transformed with pLNP1, plnp1Δznfn, or empty vector and grown to early log phase at 25°C, fivefold serially diluted, and grown at indicated temperatures. (C) Expression of lnp1Δznfn results in rescue of lnp1Δ rtn1Δ NPC aggregation. lnp1Δ rtn1Δ NIC96-GFP mutants were transformed with pLNP1, plnp1Δznfn, or empty vector and grown to early log phase at 25°C and imaged. Scale bar, 5 μm. (D) The aggregation indexes of Nic96-GFP–expressing cells. Error bars represent SE. Asterisk denotes statistical significance (p < 0.01). (E) Expression of lnp1Δznfn is not sufficient to rescue lnp1Δ defects in ER. lnp1Δ SEC61-GFP mutants were transformed with pLNP1, plnp1Δznfn, or empty vector and grown to early log phase at 25°C and imaged. Scale bar, 5 μm. (F) Percentages of cells with regions of collapsed cortical ER as indicated by lack of peripheral Sec61-GFP staining were quantified from images of Sec61-GFP–expressing cells. Error bars represent SE. Asterisk denotes statistical significance (p < 0.01).