• Journal of the Korean Society of Food Science and Nutrition
• /
• v.26 no.6
• /
• pp.1246-1251
• /
• 1997

When the membrane protein fraction of mating type a cells of heterobasidiomycetous yeast R. toruloides was phosphorylated in vitro, two phosphorylated proteins of 72Kd and 57Kd were detected on SDS-polyacryamide gel. The phosphorylation reaction was inhibited by rhodotorucine A(Rh. A) which is a sexual pheromone secreted by mating type A cells. The inhibition of phosphorylation by Rh. A was dependent on $Ca^{2+}$, and independent on $Mg^{2+}$ or calmodulin. When adding trigger peptidase(TPase) inhibitor, antipain, no inhibition of phosphory was observed. Also, by adding the trysin-digested product of Rh. A, the phosphorylation was inhibited as the action of Rh. A. From these results, it is expected that the inhibition of membrane protein phosphorylation should be caused by the digested product of Rh. A with TPase.

• Microbiology and Biotechnology Letters
• /
• v.24 no.6
• /
• pp.641-646
• /
• 1996

[$\gamma$-$^{32}$P]ATP was used to test phosphorylation of membrane proteins of mating type a cells of heterobasidiomycetous yeast Rhodosporidium toruloides separated by non-denaturing electrophoresis. The phosphoprotein was observed in the membrane proteins. The phosphorylation was inhibited by the pheromone rhodotorucine A (Rh. A) secreted by mating type A of the yeast. Rh. A didn't inhibit the phosphorylation in the presence of a trigger peptidase (TPase) inhibitor, antipain. Partially digested Rh. A by trypsin maintained the phosphorylation inhibitory activity. These results show that TPase activity plays an important role in the transduction of pheromone signal in the yeast.

• Journal of Life Science
• /
• v.7 no.4
• /
• pp.322-328
• /
• 1997

Two phosphorylated proteins having molecular weights of 57kD and 72kD were detected from the slubilized membrane protein fraction of mating type a cells of Rhodosporidium toruloides which belongs to heterobasidiomycetous yeast. The phosphorylation of the protein was inhibited by a sexual pheromone, Rhodotorucine A (Rh. A), which is secreted from mating type a cells. On the other hand, counterpart mating type A cells and M-39 strain which is a styerile mutant derived from a cells, had also the same two phosphorylated proteins, However, the phosphorylation of the protein from A cells, and M-39 strain were not inhibited by the Rh. A. It suggests that inhibition of the phosphorylation reaction by the Rh. A in mating type a cells is a mating-type-specific reaction that relate to transduction mechanism of sexual pheromone signaling.

• Microbiology and Biotechnology Letters
• /
• v.23 no.3
• /
• pp.249-254
• /
• 1995

When mating type A and a cells of heterobasidiomycetous yeast Rhodosporidium toruloides were mix-cultured, both of the mating type cells have shown strong agglutination. But this agglutination was not detactable when the A and a cell were cultured separately. From reagglutination made just after the result of disassembling the agglutination by sonication, we knew that the agglutination was sexual-agglutination, not simple physical cell agglutination. The sexual agglutination was progressed actively on logarithmic phase and, in addition, progressed faster on mating type a cell treated with rhodotorucine A. These sexual agglutination have been inhibited by several protease such as trypsin, pronase, chymotrpysin and thermolysin and inhibited by 5 mM DTT as well.

• Journal of Life Science
• /
• v.6 no.3
• /
• pp.165-170
• /
• 1996

The action of phospholipid on the rhodotorucine A(RH.A) acceptance by heterbasidiomyceteous yeast Rhodosporidium toruloides mating type a cells and the effect of medium composition during sexual differentiation were investigated. Activation of trigger peptidase(TPase)was very sensitive to the originated phospholipid from R. toruloides and was more sensitive to phospholipid liposome made up of phospholipi. Phospholopod present on the membrance of mating type a cells consists of phospatidylglycerol(PG), phosphatidylethanolamine(PE), phospatidylcholine(PC), phospatidylinositol(PI), and phosphatidylserine(PS) of 12.9, suprisingly 45.4, 11.0, and 13.9%, respectively. As the result of using C-1 and N-1 mediums which limited C and N sources capable of inhibiting the synthesis of phospholipid, it resulted inhibiting sexual dlfferentiation and production of Rh.A from mating type Acells.