FIGURE 3:
PKA modulation is sufficient but not necessary for regulation of Msn2 nuclear localization. (A–C) Kymographs of Y3630 (MSN2-GFP msn4∆ prototroph) cells immobilized in a microfluidic chamber switched from SD medium to 0% glucose at time 0 (A) or from SD lacking glucose to SD + 2% glucose at time 0 (B), or subjected to 2-min cycles between 0 and 2% glucose (C). (D–F) Kymographs of Y3817 (pka-as MSN2-GFP msn4∆) cells immobilized as previously and switched from SD medium to SD + 1 μM inhibitor 1NM-PP1 at time 0 (D) or from SD +1 M 1NM-PP1 to SD at time 0 (E) or subjected to 2-min cycles of SD and SD + 1 M 1NM-PP1 (F). (G–I) Kymographs of Y3845 (pka-wimpD139H MSN2-GFP msn4∆) cells treated as in A–C. (J–L) Kymographs of Y3964 (pka-wimpD139H snf1Δ MSN2-GFP msn4∆) cells treated as in A–C. Note that Msn2-GFP resides in the nucleus at time 0 in G and J due to the low levels of PKA activity in these strains. See also Supplemental Figure S3. Data were collected at 20-s intervals, except for A and G, which were collected at 1-min intervals.