Fig. 6

Distruction of two most likely NESs does not influence Irr1 cellular distribution upon nitrogen starvation or α-factor arrest. Cells expressing GFP-tagged Irr1 variants with one or two potential NES motifs disrupted, as indicated on the left, were grown in YPD medium and subjected to nitrogen starvation for 22 h (SD-N) or treated with α-factor for 3 h (α-factor). DAPI-DNA stained with DAPI, DIC—transmitted light. Consensus sequence for Crm1-dependent nuclear export is WX2–3WX2–3WXW, where W represents L, I, V, F or M and X—any amino acid. NES signals disrupted by site-directed mutagenesis. irr1(V248E) carries inactivated NES between positions 238–248, irr1(F986A)—between positions 982–992, irr1(V248E, F986A) has both signals inactivated