Fig 1.

(A) P. brasiliensis TPK2 complements the growth defect of the S. cerevisiae ΔTPK2 Ts mutant strain SGY446. The S. cerevisiae haploid strain SGY446 that cannot grow at 37°C was transformed with p426MET25 (a), p426MET25-GFP (b), and the p426MET25-GFP constructs for the expression of GFP-fusion proteins of the N-terminal domain (PbTpk2^(1–225)) (c), C-terminal domain (PbTpk2^(226–583)) (d)) and full-length (PbTpk2^(1–583)) PbTpk2 (e) from P. brasiliensis and incubated at 25°C and 37°C. The cells expressing the C-terminal domain (d) and full-length PbTpk2 (e) were able to grow at 37°C. (B) P. brasiliensis PbTPK2 complements the defect in the ability of the S. cerevisiae ΔTPK2 XPY5a/α strain to form pseudohyphae. The S. cerevisiae diploid strain XPY5a/α was transformed with constructs for the expression of GFP (GFP), the the N-terminal domain (PbTpk2^(1–225)), C-terminal domain (PbTpk2^(226–583) and PbTpk2^(265–583)), full-length (PbTpk2^(1–583)) PbTpk2 and the K301R full-length PbTpk2 derivative from P. brasiliensis and the transformants were analysed for pseudohyphal growth in SLAD agar, containing 50 μM (upper panel) or 200 μM (lower panel) ammonium sulfate. Single colonies from the agar plate were observed at 20x magnification in an Eclipse E-400 microscope. The scale bar is 50 μm. The cells expressing the C-terminal domains and full-length PbTpk2 were able to form pseudohyphae, but cells expressing the full-length K301R PbTpk2 derivative, defective in kinase activity, were unable to form pseudohyphae.