Mitochondrially-targeted bacterial phosphatidylethanolamine methyltransferase sustained phosphatidylcholine synthesis of a Saccharomyces cerevisiae Δpem1 Δpem2 double mutant without exogenous choline supply.
Mitochondrially-targeted bacterial phosphatidylethanolamine methyltransferase sustained phosphatidylcholine synthesis of a Saccharomyces cerevisiae Δpem1 Δpem2 double mutant without exogenous choline supply.
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线粒体靶向细菌磷脂酰乙醇胺甲基转移酶在没有外源胆碱供应的情况下维持酿酒酵母 Δpem1 Δpem2 双突变体的磷脂酰胆碱合成。
DOI：
10.1016/j.bbalip.2014.05.003
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
A. Ohta
中科院分区：
文献类型：
--
作者：
Shingo Kobayashi;Aya Mizuike;H. Horiuchi;R. Fukuda;A. Ohta
在真核细胞中，磷脂仅在针对每种磷脂种类的特定细胞器中合成。为了解释磷脂酰胆碱 (PC) 合成中这种区室特异性的原因，我们构建并表征了一种酿酒酵母菌株，该菌株缺乏内源性磷脂酰乙醇胺 (PE) 甲基转移酶，但具有来自醋杆菌的重组 PE 甲基转移酶，该酶与来自酵母 Pet100p 的线粒体靶向信号和 3 × HA 表位标签融合。我们将这种融合蛋白命名为mitopmt，通过荧光显微镜和亚细胞分级分离确定其定位于线粒体。 mitopmt的表达抑制了PEM1和PEM2双缺失突变体(pem1Δpem2Δ)的胆碱营养缺陷，使其能够在缺乏胆碱的情况下合成PC。即使肯尼迪途径因编码CTP：磷酸胆碱胞苷酰转移酶的抑制而失活，也观察到这种生长抑制，这表明在线粒体中合成的PC分布到其他细胞器而不通过挽救途径。编码线粒体磷脂酰丝氨酸脱羧酶的 PSD1 删除的 pem1Δpem2Δ 菌株能够生长，因为在乙醇胺存在下表达了 mitopmt，这意味着来自其他细胞器（可能来自 ER）的 PE 被 mitopmt 转化为 PC。这些结果表明 PC 可以移出线粒体，并提出其运动不受严格方向限制的可能性。
In eukaryotic cells, phospholipids are synthesized exclusively in the defined organelles specific for each phospholipid species. To explain the reason for this compartmental specificity in the case of phosphatidylcholine (PC) synthesis, we constructed and characterized a Saccharomyces cerevisiae strain that lacked endogenous phosphatidylethanolamine (PE) methyltransferases but had a recombinant PE methyltransferase from Acetobacteraceti, which was fused with a mitochondrial targeting signal from yeast Pet100p and a 3 × HA epitope tag. This fusion protein, which we named as mitopmt, was determined to be localized to the mitochondria by fluorescence microscopy and subcellular fractionation. The expression of mitopmt suppressed the choline auxotrophy of a double deletion mutant ofPEM1andPEM2(pem1Δpem2Δ) and enabled it to synthesize PC in the absence of choline. This growth suppression was observed even if the Kennedy pathway was inactivated by the repression ofPCT1encoding CTP:phosphocholine cytidylyltransferase, suggesting that PC synthesized in the mitochondria is distributed to other organelles without going through the salvage pathway. Thepem1Δpem2Δ strain deleted forPSD1encoding the mitochondrial phosphatidylserine decarboxylase was able to grow because of the expression of mitopmt in the presence of ethanolamine, implying that PE from other organelles, probably from the ER, was converted to PC by mitopmt. These results suggest that PC could move out of the mitochondria, and raise the possibility that its movement is not under strict directional limitations.
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膜相关酶磷脂酰丝氨酸合酶的酵母结构基因的分离。
DOI：
10.1073/pnas.80.23.7279
发表时间：
1983
影响因子：
11.1
作者：
Letts,VA;Klig,LS;Bae-Lee,M;Carman,GM;Henry,SA
通讯作者：
Henry,SA
影响因子：
3.9
作者：
Forsha, D;Church, C;Poyton, RO
通讯作者：
Poyton, RO
酿酒酵母中磷脂酰丝氨酸的合成。
DOI：
--
发表时间：
1984
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Bae-Lee,MS;Carman,GM
通讯作者：
Carman,GM
通过与大肠杆菌突变体互补，从酿酒酵母中克隆编码磷脂酰丝氨酸脱羧酶的基因 (PSD1)。
DOI：
--
发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Clancey,CJ;Chang,SC;Dowhan,W
通讯作者：
Dowhan,W
DOI：
10.1073/pnas.95.1.150
发表时间：
1998-01-06
影响因子：
11.1
作者：
Mandala, SM;Thornton, R;Spiegel, S
通讯作者：
Spiegel, S