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首页> 外文期刊>Molecular and Cellular Biology >The membrane-associated enzyme phosphatidylserine synthase is regulated at the level of mRNA abundance.
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The membrane-associated enzyme phosphatidylserine synthase is regulated at the level of mRNA abundance.

机译：膜相关的酶磷脂酰丝氨酸合酶在mRNA丰度水平受到调节。

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To precisely define the functional sequence of the CHO1 gene from Saccharomyces cerevisiae, encoding the regulated membrane-associated enzyme phosphatidylserine synthase (PSS), we subcloned the original 4.5-kilobase (kb) CHO1 clone. In this report a 2.8-kb subclone was shown to complement the ethanolamine-choline auxotrophy and to repair the defect in the synthesis of phosphatidylserine, both of which are characteristic of cho1 mutants. When this subclone was used as a hybridization probe of Northern and slot blots of RNA from wild-type cells, the abundance of a 1.2-kb RNA changed in response to alterations in the levels of the soluble phospholipid precursors inositol and choline. The addition of inositol led to a 40% repression of the 1.2-kb RNA level, while the addition of choline and inositol led to an 85% repression. Choline alone had little repressive effect. The level of 1.2-kb RNA closely paralleled the level of PSS activity found in the same cells as determined by enzyme assays. Disruption of the CHO1 gene resulted in the simultaneous disappearance of 1.2-kb RNA and PSS activity. Cells bearing the ino2 or ino4 regulatory mutations, which exhibit constitutively repressed levels of a number of phospholipid biosynthetic enzymes, had constitutively repressed levels of 1.2-kb RNA and PSS activity. Another regulatory mutation, opi1, which causes the constitutive derepression of PSS and other phospholipid biosynthetic enzymes, caused the constitutive derepression of the 1.2-kb RNA. When cho1 mutant cells were transformed with the 2.8-kb subclone on a single-copy plasmid, the 1.2-kb RNA and PSS activity levels were regulated in a wild-type fashion. The presence of the 2.8-kb subclone on a multicopy plasmid, however, led to the constitutive overproduction of 1.2-kb RNA and PSS in cho1 cells.
机译：为了精确地定义来自酿酒酵母的CHO1基因的功能序列，该膜编码受调节的膜相关酶磷脂酰丝氨酸合酶（PSS），我们亚克隆了原始的4.5千碱基（kb）CHO1克隆。在该报告中，显示了一个2.8kb的亚克隆，可补充乙醇胺-胆碱的营养缺陷并修复磷脂酰丝氨酸合成中的缺陷，这两者都是cho1突变体的特征。当该亚克隆用作来自野生型细胞的RNA的Northern杂交和狭缝印迹的杂交探针时，响应于可溶性磷脂前体肌醇和胆碱水平的变化，1.2kb RNA的丰度发生了变化。肌醇的添加导致1.2-kb RNA水平的40％抑制，而胆碱和肌醇的添加导致85％的抑制。单独的胆碱抑制作用很小。通过酶法测定，1.2 kb RNA的水平与在同一细胞中发现的PSS活性水平非常接近。 CHO1基因的破坏导致1.2-kb RNA和PSS活性同时消失。具有ino2或ino4调节突变的细胞，其组成性抑制水平的多种磷脂生物合成酶，其组成性抑制水平的1.2-kb RNA和PSS活性。另一个调节性突变opi1引起PSS和其他磷脂生物合成酶的组成型抑制，导致1.2kb RNA的组成型抑制。当在单拷贝质粒上用2.8kb亚克隆转化cho1突变细胞时，以野生型方式调节1.2kb RNA和PSS活性水平。然而，多拷贝质粒上2.8kb亚克隆的存在导致cho1细胞中1.2kb RNA和PSS的组成型过量生产。

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