| Co-Investigator(Kenkyū-buntansha) |
HARUMOTO Terue Nara Women's University, Graduate School of Humanities and Sciences, Associate Professor, 大学院・人間文化研究科, 助教授 (80198936)
USUKI Yoshinosuke Osaka City University, Graduate School of Science, Lecturer, 大学院・理学研究科, 講師 (30244651)
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| Budget Amount *help |
¥12,800,000 (Direct Cost: ¥12,800,000)
Fiscal Year 2002: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2001: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2000: ¥6,400,000 (Direct Cost: ¥6,400,000)
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| Research Abstract |
1. Conjugation of ciliate Blepharisma japonicum Conjugation in Blepharisma japonicum is induced byinteraction between complementary mating-types I and II, which excrete blepharmone (gamone 1) and blepharismone (gamone 2), respectively. Gamone 1 transforms type II cells such that they can unite, and gamone 2 similarly transforms type I cells. The partial amino acid sequence of four peptides obtained from gamone I was determined. By using mRNA from mating type, gamone 1 cDNA was isolated by rapid amplification. The cDNA sequence contains an ORF of 305 amino acids and codes a novel protein. We also estimated that the sugar sequence for the glycoprotein consisted from four N-acetylglucosamines and three mannoses.
To elucidate the receptor of blepharismone, a photoaffinity label probe of blepharismone is designed. We synthesized a model compound Nbenzoyl blepharismone based on the strategy developed before. The model compbund did not have mating inducing activity as expected, it was found to
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have the significant activity ofmating inhibition.
2-1. Defense toxins of Climacostomum Virens The heterotrich ciliate Climacostomum virens has colourless cortical granules morphologically similar to pignentocysts (extrusive pigment granules) in the two coloured heterotrichs, Blepharisma japonicum and Stentor coeruleus, which are known to function as defensive organelles against predators. Climacostol, a defense toxin of the heterotrich ciliate Climacostomum virens was isolated and the structure was established as 5-(Z)-non-2-enyl-benzene-1, 3-diol. The structure was rigorously confirmed by the total synthesis. The two congeners of climacostol contained in this ciliate were determined as 5-(Z, Z, Z)-undeca-2, 5, 8-trienyl-benzene-I, 3-diol and 5-(Z, Z)-undeca-2, 5-dienyl-benzene-1, 3-diol.
2-2. Defense toxin of Spirostomum teres A heterotrich ciliate Spirostomum teres has colorless cortical granules that are mophologically similar to the pigment granules of B. japonicum and S. coeruleus, and the predatory ciliate D. margaritifer rarely attacks S. teres. We found the ethanol extracts of S. teres shown to be toxic to D. margaritifer. The structure of a toxic compound named spirostomin was elucidated as spiro-2,5 -dimethyl-5,6,7,8-tetrahydronaphtalene-1,4-dione-8,6'-pyran-2',5'-dione. Less
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