204-07-21369C Petersen and Hagan, Supplementary Figure 1

1. 25ºC  37 ºC 25ºC  37 ºC 204-07-21369C Petersen and Hagan, Supplementary Figure 1 A C D M WCE wt wt sty1::ura4+ -P-S402 116.2 Cdc2 97.4 78.0 Plo1 55.0 Cdc2 42.0 0 30 60 90 120 150 180 180 0 60 0 30 60 90 120 150 Time (min) Time (min) B E Sorbitol 1M Heat 37C H2O2 25 mM 0 15 60 15 60 0 15 60 -P-S402 + -Plo1 -P-S402 -Plo1 -P-S402 -tubulin Cdc2

2. 25ºC  37 ºC 204-07-21369C Petersen and Hagan, Supplementary Figure 2 B A 25ºC  37 ºC 40 Asynch. cdc25.22 25ºC  37 ºC 30 0 30 60 90 120 150 180 % Septation 20  P-S402 10 Cdc2 0 120 150 180 210 240 0 30 60 90 Time (min)  P-S402 cdc2 0 60 120 180 240 C 0 10 20 30 45 60 90 120 150 180 -P-TY -P-Y Sty1.myc -AB D 0 30 60 90 120 150 180 37°C Plo1 -myc I.P’s Sty1. myc Plo1 (Input) -AB E 0 30 60 90 120 150 180 37°C Plo1 -myc I.P’s Sty1. myc Plo1 (Input)

3. 204-07-21369C Petersen and Hagan, Supplementary Figure 3 Before centrifugation After centrifugation

4. 204-07-21369C Petersen and Hagan, Supplementary Figure 4 I.P. of HA-Plo1 after Heatshift Total 0 10 20 30 60 90 M Anti HA Total M 0 10 20 30 60 90 Anti Plo1

5. + 204-07-21369C Petersen and Hagan, Supplementary Figure 5 B Recovery from stress A Mitotic commitment Heat / Centrifugation SPB nuclear envelope Spc1/Sty1 phosphate P Actin Response to deal with the stress incl. stop division, de-polarise actin G2 M ? Return to normal growth P P Cdc25 Cdc2 Cdc2 P Plo1 CyclinB CyclinB Wee1 ? Re-entry into cycle Re-initiation of tip growth Plo1 P Spc1/Sty1 Cut12 Re-organisation of actin at the cell tips

6. Petersen and Hagan, Supplementary Table 1 Strains used in this study IH163 h- 972 Lab stock IH367 h- ura4.d18 leu1.32his2 Lab stock IH634 h+ cdc25.22 ura4.d18 leu1-his- Lab stock IH635 h- cdc25.22 ura4.d18 Lab stock IH565 h+ cdc25.22 ura4.d18 leu1Lab stock IH666 h+ cut12.s11 leu1.32 Bridge et al. 1998 IH740 h+ cdc25.22cut12.s11 ura4.d18 leu1.32 Bridge et al. 1998 !H1314 h- leu1::pint5(ura4+)- plo1.NHA ura4.d18 Tanaka et al. 2001 IH1624 h+ plo1.ts2ura4.d18 leu1.32his2 ade6.M210MacIver et al. 2003 IH2420 h- sty1::ura4+Millar et al. 1995 IH2783 h- wis1.DD::ura4+ ura4.d18 leu1.32Shiozaki et al. 1998 IH2873 h+ wis1::ura4 ura4.d18 Millar et al. 1995 IH2986 h90atf1::ura4 ura4.d18 leu1.32 Quinn et al. 2002 IH3124 h+ plo1.S402A ura4.d18 leu1.32his2This study IH3125 h+ plo1.S402E ura4.d18 leu1.32his2 This study IH3224 h+ cdc25.22 wis1.DD::ura4+ leu1.32his2 This study IH3225 h+ cdc25.22 cut12.s11 plo1.S402A ura4.d18 leu1.32his2This study IH3226 h- cdc25.22 cut12.s11 plo1S402E ura4.d18 leu1.32 This study IH3270 h+ cdc25.22 wis1,DD plo1.S402A ura4.d18 leu1.32his2This study IH3272 h+ cdc25.22 wis1.DD plo1.S402Eura4.d18 leu1.32his2This study IH3275 h- cdc25.22 plo1.S402Aura4.d18 leu1.32This study IH3277 h- cdc25.22 plo1.S402Eura4.d18 leu1.32This study IH3278 h- wis1.DD plo1.S402Aura4.d18 leu1.32This study IH3279 h+ wis1.DD plo1.S402Eura4.d18 leu1.32his2This study IH3282 h- cut12.s11 plo1.S402A ura4.d18 leu1.32 ade6This study IH3283 h- cut12.s11 plo1.S402E ura4.d18 leu1.32 ade6This study IH3286 h+ sty1::ura4+ plo1.S402A ura4.d18 leu1.32 his2 This study IH3287 h+ sty1::ura4+ plo1.S402E ura4.d18 leu1.32 his2This study IH3306 h- cdc25.22 leu1::plo1.S402con:ura4+ ura4.d18 This study IH3307 h- cdc25.22 leu1::plo1.S402Acon:ura4+ ura4.d18 This study IH3308 h- cdc25.22 leu1::plo1.S402Econ:ura4+ ura4.d18 This study IH3380 h+ wis1.DD::ura4+ plo1.ts2 leu1.32 his2 ura4.d18 This study IH3724 h-plo1.ts2 sty1::ura4 This study IH3758 h-plo1.S402A This study IH3759 h- plo1.S402E This study IH3789 h- spc1-12 myc (ura4+)  Gaits et al. 1995 IH3988 h- spc1-12 myc (ura4+) leu1.32 p81NPkPlo1  This study References Bridge, A. J., Morphew, M., Bartlett, R., and Hagan, I. M. (1998). The fission yeast SPB component Cut12 links bipolar spindle formation to mitotic control. Genes and Development 12, 927-942. Gaits F, Degols G, Shiozaki K, Russell P. (1998) Phosphorylation and association with the transcription factor Atf1 regulate localization of Spc1/Sty1 stress-activated kinase in fission yeast. Genes Dev. 12, 1464-73. MacIver, F. H., Tanaka, K., Robertson, A. M., and Hagan, I. M. (2003). Physical and functional interactions between polo kinase and the spindle pole component Cut12 regulate mitotic commitment in S. pombe. Genes Dev 17, 1507-1523. Millar, J. B., Buck, V., and Wilkinson, M. G. (1995). Pyp1 and Pyp2 PTPases dephosphorylate an osmosensing MAP kinase controlling cell size at division in fission yeast. Genes Dev 9, 2117-2130. Quinn, J., Findlay, V. J., Dawson, K., Millar, J. B., Jones, N., Morgan, B. A., and Toone, W. M. (2002). Distinct regulatory proteins control the graded transcriptional response to increasing H2O2 levels in fission yeast Schizosaccharomyces pombe. Mol Biol Cell 13, 805-816. Shiozaki, K., Shiozaki, M., and Russell, P. (1998). Heat stress activates fission yeast Spc1/StyI MAPK by a MEKK-independent mechanism. Mol Biol Cell 9, 1339-1349 Tanaka, K., Petersen, J., MacIver, F., Mulvihill, D. P., Glover, D.M. and Hagan, I.M. (2001). The role of Plo1 kinase in mitotic commitment and septation in Schizosaccharomyces pombe. EMBO J. 20 1259-1270