| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1995: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1994: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1993: ¥900,000 (Direct Cost: ¥900,000)
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| Research Abstract |
1.Mitochondria and mitochondrial nucleoids (mt-nucleoids) of the yeast, Saccharomyces cerevisiae were vitally-stained with two fluorochromes, 3,3'-dihexyloxacarbocyanine iodide [DiOC_<6(>3)] and 4', 6-diamidino-2-phenylindole (DAPI), respectively in the different types of cells during the life cycle. The results showed that the fusion and fragmentation of mitochondria occurred frequently during vegetative growth, sporulation, mating and germination of yeasts.
2.Mt-nucleoids isolated from the yeast Saccharomyces cerevisiae were analyzed to indentify the protein components that are involved in the compact packaging of mtDNA.DNA-binding proteins with molecular masses of 67 kDa, 52 kDa, 50 kDa, 38 kDa 30 kDa and 20 kDa were separated from isolated mt-nucleoids. The fraction that contained the DNA-binding proteins was able to fold mtDNA compactly into nucleoid-like structures. These results suggested that at least six different DNA-binding proteins were involved in the organization of mt-nucleoids. An antibody was raised against 67-kDa mt-nucleoid protein of S.cerevisiae mt-nucleoids.
3.Mt-nucleoids were isolated at high purity from the yeast Pichia jadinii and other four species of yeasts that belong to the genus Pichia, Williopsis. SDS-polyacryl amide gel electrophoresis and immunoblotting suggested that the diversity of mt-nucleoid protein occurred in yeasts.
4.Relationships between the morphology of mt-nucleoids and distribution of actin were investigated by DAPI-staining and by the staining with Rhodamine-phalloidin. During sporulation of diploid cells, mt-nucleoids formed a network of fluorescent strings and actin dots were frequently overlapped on the mt-nucleoids.
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