1. 204-07-21369C Petersen and Hagan, Supplementary Figure 1 Cdc2 Plo1  -P-S402 0306090120150180 wt Time (min) C 0600306090120150 180 wtsty1::ura4 + Time (min) D 25ºC  37 ºC  -P-S402  -tubulin Cdc2 015 60 Sorbitol 1M Heat 37  C 015 60 H2O2H2O2 25 mM E 116.2 97.4 78.0 55.0 42.0 A 25ºC  37 ºC WCEM  -P-S402  -Plo1  -P-S402 +  -Plo1 B

2. Asynch. cdc25.22 25ºC  37 ºC B 0306090120150180 Cdc2  P-S402 25ºC  37 ºC 0 10 20 30 40 0 306090 120150180210240 0 60 120 180 240 cdc2 Time (min) % Septation A  -P-Y  -P-TY 0 10 20 30 45 60 90 120 150 180 Sty1.myc 25ºC  37 ºC C 0 30 60 90 120 150 180 -AB 37°C Plo1 Sty1. myc Plo1 (Input)  -myc I.P’s 0 30 60 90 120 150 180 -AB 37°C Plo1 Sty1. myc Plo1 (Input)  -myc I.P’s D E 204-07-21369C Petersen and Hagan, Supplementary Figure 2

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

4. I.P. of HA-Plo1 after Heatshift Total0 10 20 30 60 90 Anti HA 0 10 20 30 60 90 Anti Plo1 Total M M 204-07-21369C Petersen and Hagan, Supplementary Figure 4

5. CyclinB Cdc2 Cdc25 Wee1 PP Plo1 Spc1/Sty1 Mitotic commitment + Cut12 G2M P Re-initiation of tip growth Plo1 Spc1/Sty1 Recovery from stress P Re-entry into cycle Re-organisation of actin at the cell tips nuclear envelope SPB P phosphate Actin Heat / Centrifugation Response to deal with the stress incl. stop division, de-polarise actin Return to normal growth CyclinB Cdc2 A B ? ? 204-07-21369C Petersen and Hagan, Supplementary Figure 5

6. Strains used in this study IH163 h - 972 Lab stock IH367h - ura4.d18 leu1.32 his2Lab stock IH634 h + cdc25.22 ura4.d18 leu1- his- Lab stock IH635h - cdc25.22 ura4.d18Lab stock IH565h + cdc25.22 ura4.d18 leu1Lab stock IH666 h + cut12.s11 leu1.32Bridge et al. 1998 IH740 h + cdc25.22 cut12.s11 ura4.d18 leu1.32Bridge et al. 1998 !H1314 h - leu1::pint5(ura4 + )- plo1.NHA ura4.d18Tanaka et al. 2001 IH1624 h + plo1.ts2 ura4.d18 leu1.32 his2 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.d18Millar et al. 1995 IH2986 h 90 atf1::ura4 ura4.d18 leu1.32Quinn et al. 2002 IH3124 h + plo1.S402A ura4.d18 leu1.32 his2This study IH3125 h + plo1.S402E ura4.d18 leu1.32 his2This study IH3224 h + cdc25.22 wis1.DD::ura4 + leu1.32 his2This study IH3225 h + cdc25.22 cut12.s11 plo1.S402A ura4.d18 leu1.32 his2This study IH3226 h - cdc25.22 cut12.s11 plo1S402E ura4.d18 leu1.32This study IH3270 h + cdc25.22 wis1,DD plo1.S402A ura4.d18 leu1.32 his2This study IH3272 h + cdc25.22 wis1.DD plo1.S402E ura4.d18 leu1.32 his2This study IH3275 h - cdc25.22 plo1.S402A ura4.d18 leu1.32This study IH3277 h - cdc25.22 plo1.S402E ura4.d18 leu1.32This study IH3278 h - wis1.DD plo1.S402A ura4.d18 leu1.32This study IH3279 h + wis1.DD plo1.S402E ura4.d18 leu1.32 his2This 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 his2This study IH3287 h + sty1::ura4 + plo1.S402E ura4.d18 leu1.32 his2This study IH3306h - cdc25.22 leu1::plo1.S402con:ura4 + ura4.d18This study IH3307h - cdc25.22 leu1::plo1.S402Acon:ura4 + ura4.d18This study IH3308h - cdc25.22 leu1::plo1.S402Econ:ura4 + ura4.d18This study IH3380h + wis1.DD::ura4 + plo1.ts2 leu1.32 his2 ura4.d18This study IH3724h- plo1.ts2 sty1::ura4 This study IH3758h- plo1.S402A This study IH3759h- 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 Petersen and Hagan, Supplementary Table 1 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 H 2 O 2 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