| Project/Area Number |
60480466
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Biological pharmacy
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| Research Institution | National Institute of Health, Japan |
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Principal Investigator |
AKAMATSU Yuzuru Director, Dept. of Chemistry, N.I.H., Japan, 化学部, 部長 (00072900)
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| Co-Investigator(Kenkyū-buntansha) |
KUGE Osamu Senior Investigator, Dept. of Chemistry, N.I.H., Japan, 化学部, 研究員 (30177977)
AMANO Fumio Chief Investigator, Dept. of Chemistry, N.I.H., Japan, 化学部, 主任研究員 (90142132)
TANAKA Yasuhito Chief Investigator, Dept. of Chemistry, H.L.H., Japan, 化学部, 主任研究員 (30113484)
KITAGAWA takayuki Section Chief, Dept. of Chemistry, N.I.H., Japan, 化学部, 室長 (80092188)
NISHIJIMA Masahiro Section Chief, %dept. of Chemistry, N.I.H., Japan, 化学部, 室長 (60072956)
久世 洋子 国立予防衛生研究所, 化学部, 研究員 (50178000)
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| Project Period (FY) |
1985 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1986: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1985: ¥4,200,000 (Direct Cost: ¥4,200,000)
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| Keywords | mammalian cell variant / CHO cell / enveloped RNA virus / macrophage / phospholipid-base exchange enzyme / phosphatidylserine / phospholipid metabolism / ホスホリパーゼA_2 / RAW264細胞株 / 脂質塩基交換反応 / ホスファチジルセリン合成系 / ホスホリパーゼ |
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| Research Abstract |
The aims of this research are to establish variants defective in the phospholipid metabolisms in cultured mammalian cells and, by suing these variants, to clarify the mechanisms of the phospholipid biosynthesis and its regulation and finally to explore the roles of phospholipids in biosynthesis and functions of biological membranes.
1. We established the variants lacking biosynthesis of phosphatidylserine (PS) from Chines hamster ovary (CHO) cells, and biosynthesis of PS was shown catalysed by the phospholipid-choline exchange enzymes and the importance of PS in cell growth was also pointed out.
2. By obtaining PS auxotroph (PSA 1-3) the existence of two serine exchange enzymes (Enzyme I and Enzyme II) in mammalian cells was found out for the first time. Furthermore, the defect in Enzyme I was shown to induce the decrease in PS content and the consequent death of the cells.
3. By analyzing the infection of this variant cells with enveloped RNA virus the functions of PS in this process was clarified. E expect this study gives important informations for finding the procedures to decrease virus maturation.
4. The variant defective in the regulatory mechanism of PS biosynthesis was also isolated. In the parent strain PS synthesis was suppressed specifically with PS added to the growth medium. In the variant cells, however, this suppression was lacking. Further investigations are now in progress.
5. A gene from CHO cells which complement PS requirement in PSA-3 cells was isolated and the relationship between this gene and the PS biosynthesis was established.
6. Variants from macrophage cell line defective in phospholipase A_2 activity were obtained and the existence of at least two phospholipase A_2 was suggested.
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