Lte1 influences localization of Bfa1-GFP at SPBs. (A) Depletion of Lte1 arrests growth in strains expressing Bfa1-GFP. MGY252 (lte1Δ BFA1-GFP YCplac33 URA3-LTE1) and MGY260 (lte1Δ BFA1-GFP GAL1_pr–lte1-F1387E–3HA) were tested for growth at 30°C after spotting on YEP galactose (GAL) and YEP glucose (GLU) agar to induce or repress expression of Lte1-F1387E, respectively. (B) BFA1-GFP is colethal with lte1Δ and can be rescued by activation of the MEN. MGY252 (lte1Δ BFA1-GFP YCplac33 URA3-LTE) was transformed with dbf2 HyA or deleted for KIN4 and spotted onto YEP dextrose (YPD) and 5-fluoroorotic acid (5FOA) agar to test for growth at 30°C in the absence of Lte1. (C) lte1Δ BFA1-GFP cells arrest in anaphase with Bfa1-GFP at both SPBs. MGY260 lte1Δ expresses Bfa1-GFP (red) and Tub1-CFP (green) from their endogenous promoters and expresses Lte1-F1387E from the GAL1 promoter of the integrative plasmid pMG75. +Lte1 indicates Lte1 expressed during growth in galactose-containing medium; −Lte1 indicates Lte1 expression repressed by 4–6-h growth with glucose. Expression of Bfa1-GFP and Tub1-CFP was also monitored in MGY272 (lte1Δ BFA1-GFP TUB1-CFP), which is kept alive by pDbf2-1c expressing Dbf2 HyA, and in MMY41 (lte1Δ kin4Δ BFA1-GFP TUB1-CFP). Both strains were cultivated at 30°C. (D) Bfa1 (green) and DNA (blue) were visualized in MGY274 (mob1-77 BFA1-GFP) cells arrested in anaphase by incubation at 37°C for 2.5 h. (E) The percentage of cells expressing Bfa1-GFP at one or two SPBs in late anaphase cells is presented in the yeast strains indicated (n > 100). Bars, 5 µm.