grisea

(hypothetical protein), N. crassa (PLA2), C. globosum (hypothetical protein), P. anserina (hypothetical protein) and G. zeae (PLA2). The alignment was done using MCOFFEE and visualized using the program GeneDoc. Only the catalytic core of these proteins is shown in this alignment, from amino acids 192 to 611 (in reference to the multiple alignment position). The black shading with white letters indicates 100% identity, gray shading with white letters indicates 75–99% identity, gray shading with black letters indicates 50–74% identity. Effects of PLA2 effectors on the yeast to mycelium transition and the yeast cell cycle S. schenckii is not a genetically manageable organism, therefore, effectors of PLA2 were tested for their CHIR-99021 research buy effects on the yeast to mycelium transition and the yeast cell cycle. Arachidonic acid is the primary product of cPLA2 activity on phospholipids, while AACOCF3 and isotetrandrine are inhibitors

of PLA2 activity. AACOCF3 is a known competitive inhibitor of PLA2 [46]. It is an analogue of arachidonic acid and presumably binds directly to the active site of the enzyme. AZD8931 research buy It is a potent and selective inhibitor of cytosolic phospholipase A [46]. Isotetrandrine on the other hand is an alkaloid that has been reported to interfere with G protein activation of PLA2 [47]. Figure 6 shows the percentage of yeast cells forming germ tubes in the presence and absence of arachidonic acid, AACOCF3 and isotetrandrine. This figure shows that these latter compounds significantly stimulated the yeast to mycelium transition at 6 and 9 h of incubation when the control cells are in the process of DNA synthesis and germ tube emergence [2]. The percent stimulation was approximately 68% and 33% at 6 h and 9 h of incubation in the presence of both AACOCF3 and isotetrandrine. In the presence of arachidonic acid a slight

(25%) non-significant inhibition was observed at 6 h of incubation. The degree of stimulation caused by the addition of AACOCF3 and isotetrandrine was similar even though the mechanism of action of these compounds is completely different. Figure 6 Effects of SSPLA 2 effectors on the yeast to mycelium transition. Yeast cells grown, harvested, synchronized and selected by filtration as described in Methods were induced to Gemcitabine cost form germ tubes in a basal medium with glucose at pH 4.0 and incubated at 25°C in the presence and absence of arachidonic acid (40 μM), AACOCF3 (100 μM; Nonadeca-4,7,10,13-tetraenyl-trifluoro-methyl ketone)) and isotetrandrine (50 μM; 6,6′,7,12-tetra methoxy-2,2′-dimethyl-berbaman). All values are given as the average percentage ± one SD of at least three independent experiments. The Student’s t test was used to determine the statistical significance of the data at a 95% confidence level. Values that differ significantly from those of the control at 95% confidence level are marked with an asterisk.