Adaptation mechanism of membrane lipids and its genetic control

膜脂适应机制及其遗传调控

• 批准号: 61480465
• 负责人: NOZAWA Yoshinori
• 金额: $ 3.97万
• 依托单位: Gifu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-61480465/
• 关键词: membrane fluidity; low-temperature-shift; fatty acid desaturase; 膜流動性; 低温シフト; DNA; cDNAクローニング; 環境適応機構; 生体膜脂質; 遺伝的解析; 膜物性修飾

Temperature shift of Tetrahymena cells from 39 C to 15 C induced alterations in fatty acid composition of membrane phospholipids, a decrease in satureated palmitic acid and increases in unsaturated fatty acids such as -linoleinic and linoleic acids, causing its plasma membrane more fluid as inferred with ESR and freeze-fracture electron microscopy. Although low-temperature acclimation did not affect the viability of the cells, aggregation of monosome to form polysome was observed. Whether the increase in polysome content was derived from newly transcribed mRNA or not is to be clarified. Two dimensio- l gel electrophoretic analyses of microsomal proteins from low-temperature-shifted cells showed time-dependent increases in tubulin and some proteins which have lower apparent molecular masses than fatty acyl CoA-desaturase. Using two coding regions of rat liver stearyl-CoA desaturase cDNA, Southern blotting of Tetrahymena DNA was performed. Under low stringency of 35% formamide and 5xSSC, at 28 C for 24hr, EcoRI-digested DNA hybridized with one of the desaturase cDNA fragment. Furthermore, EcoRV- and BglII-digested DNA showed hybridizing bands of 2.6kb and 2.7kb, respectively, indicating a possibility of cloning Tetrahymena enzymes using rat-derived DNA as a probe.

四膜虫细胞从39℃到15℃的温度变化引起膜磷脂脂肪酸组成的变化，饱和棕榈酸的减少，以及亚油酸和亚油酸等不饱和脂肪酸的增加，导致其质膜更多的液体，如ESR和冷冻断裂电子显微镜所推断的那样。虽然低温驯化对细胞活力没有影响，但观察到单体聚集形成多聚体。多聚体含量的增加是否来自新转录的mRNA仍有待澄清。低温移位细胞微粒体蛋白质的双向L凝胶电泳法分析表明，微管蛋白和某些表观分子质量低于脂肪酰辅酶A去饱和酶的蛋白质随时间而增加。利用大鼠肝脏硬脂酰辅酶A去饱和酶基因的两个编码区，进行了四膜虫DNA的Southern杂交。在35%甲酰胺和5xSSC的低强度下，在28℃下24小时，EcoRI酶切的DNA与其中一个去饱和酶基因片段杂交。此外，EcoRV和BglII消化的DNA分别显示2.6kb和2.7kb的杂交条带，这表明以大鼠来源的DNA为探针克隆四膜虫酶是可能的。

项目成果

Nozawa,Y.: "Advances in Membrane Fluidity;Regulation of Membrane Fluidity in Unicellular Organisms" Aloia,R.C.(ed.)A,Alan R.Liss,Inc., 20 (1987)

Nozawa,Y.：“膜流动性的进展；单细胞生物膜流动性的调节”Aloia,R.C.(ed.)A,Alan R.Liss,Inc., 20 (1987)

Umeki,L.: Biol,Chem.357. 61-65 (1986)

Umeki，L.：生物，化学.357。

Nozawa,Y.: "Regulation of membrane fluidity in unicellular organisms in "Physiological regulation of membrane fluidity"" Alan R.Liss,Inc, 18 (1988)

Nozawa,Y.：“膜流动性的生理调节”中单细胞生物膜流动性的调节”Alan R.Liss,Inc, 18 (1988)

Nozawa Y.; Umeki S.: Regulation of membrane fluidity in unicellular organisms. in "Physiological regulation of membrane fluidity". Alan R Lis s, Inc, 239-257 (1988)

野泽Y.；

Arai,H.: J.Biochem.101. 1059-1067 (1987)

Arai，H.：J.Biochem.101。