Figure 8: Ste20 promotes mitotic exit parallel to Lte1.
From: Temporal and compartment-specific signals coordinate mitotic exit with spindle position
(a,b) SPOC integrity of spo12Δ kin4Δ cells upon mother cell targeting of the indicated GFP-tagged proteins through SFK1-GBP. Graph is an average of three independent experiments. Error bars show s.d. Per experiment, 100 cells were counted per cell type. Asterisk indicates significant difference from no-GFP control (two-tailed Student’s t-test, P<0.05). (c) Representative images showing Ste20-GFP localization in the absence (upper row) and presence (lower row) of Sfk1-GBP; m, mother, d, daughter cell. Scale bars: 3 μm. (d–f) Growth analysis showing serial dilutions of the indicated cell types. SPO12 or LTE1 deletion was complemented by the corresponding wild-type gene present on an URA3-based centromeric plasmid (pRS316). URA3-based plasmid loss was induced on 5FOA plates. Leucine lacking medium (SC-LEU) selects for LEU2 based-centromeric plasmids (pRS315). ste20-kd: kinase inactive version of STE20 (in d). (g) Percentage of SPOC-deficient phenotypes in STE20-GFP SFK1-GBP cells with indicated gene deletions. Graph is an average of three independent experiments. Per experiment, 100 cells were counted per cell type. Error bars show s.d. Asterisk indicates significant difference from kar9Δ STE20-GFP SFK1-GBP cells (two-tailed Student’s t-test, P<0.05). (h) Serial dilutions of indicated cell types bearing pRS316-LTE1 (LTE1 in URA3-based centromeric plasmid) were spotted on SC (nonselective) and 5FOA (negative selection for URA3-based plasmids). (i) Current model of MEN activation by compartment-specific and unspecific factors. Arrows indicate activation, whereas capped lines indicate inhibition.
