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酿酒酵母肌醇-1-磷酸合酶(INO1)基因的表达受影响磷脂合成的因子调控。

Expression of the Saccharomyces cerevisiae inositol-1-phosphate synthase (INO1) gene is regulated by factors that affect phospholipid synthesis.

作者信息

Hirsch J P, Henry S A

出版信息

Mol Cell Biol. 1986 Oct;6(10):3320-8. doi: 10.1128/mcb.6.10.3320-3328.1986.

Abstract

The INO1 gene of Saccharomyces cerevisiae encodes the regulated enzyme inositol-1-phosphate synthase, which catalyzes the first committed step in the synthesis of inositol-containing phospholipids. The expression of this gene was analyzed under conditions known to regulate phospholipid synthesis. RNA blot hybridization with a genomic clone for INO1 detected two RNA species of 1.8 and 0.6 kb. The abundance of the 1.8-kb RNA was greatly decreased when the cells were grown in the presence of the phospholipid precursor inositol, as was the enzyme activity of the synthase. Complementation analysis showed that this transcript encoded the INO1 gene product. The level of INO1 RNA was repressed 12-fold when the cells were grown in medium containing inositol, and it was repressed 33-fold when the cells were grown in the presence of inositol and choline together. The INO1 transcript was present at a very low level in cells containing mutations (ino2 and ino4) in regulatory genes unlinked to INO1 that result in inositol auxotrophy. The transcript was constitutively overproduced in cells containing a mutation (opi1) that causes constitutive expression of inositol-1-phosphate synthase and results in excretion of inositol. The expression of INO1 RNA was also examined in cells containing a mutation (cho2) affecting the synthesis of phosphatidylcholine. In contrast to what was observed in wild-type cells, growth of cho2 cells in medium containing inositol did not result in a significant decrease in INO1 RNA abundance. Inositol and choline together were required for repression of the INO1 transcript in these cells, providing evidence for a regulatory link between the synthesis of inositol- and choline-containing lipids. The level of the 0.6-kb RNA was affected, although to a lesser degree, by many of the same factors that influence INO1 expression.

摘要

酿酒酵母的INO1基因编码受调控的酶——肌醇-1-磷酸合酶,该酶催化含肌醇磷脂合成中的首个关键步骤。在已知调控磷脂合成的条件下,对该基因的表达进行了分析。用INO1的基因组克隆进行RNA印迹杂交,检测到1.8 kb和0.6 kb的两种RNA。当细胞在磷脂前体肌醇存在的情况下生长时,1.8 kb RNA的丰度以及合酶的酶活性都大幅降低。互补分析表明,该转录本编码INO1基因产物。当细胞在含肌醇的培养基中生长时,INO1 RNA水平被抑制了12倍;当细胞在肌醇和胆碱同时存在的情况下生长时,其被抑制了33倍。在与INO1不连锁的调控基因中含有导致肌醇营养缺陷型的突变(ino2和ino4)的细胞中,INO1转录本的水平非常低。在含有导致肌醇-1-磷酸合酶组成型表达并导致肌醇排泄的突变(opi1)的细胞中,该转录本组成型过量产生。还在含有影响磷脂酰胆碱合成的突变(cho2)的细胞中检测了INO1 RNA的表达。与野生型细胞中观察到的情况相反,cho2细胞在含肌醇的培养基中生长不会导致INO1 RNA丰度显著降低。在这些细胞中抑制INO1转录本需要肌醇和胆碱同时存在,这为含肌醇和含胆碱脂质的合成之间的调控联系提供了证据。0.6 kb RNA的水平也受到许多影响INO1表达的相同因素的影响,但程度较小。

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