Phospholipid Metabolism and Membrane Function

磷脂代谢和膜功能

• 批准号: 7889114
• 负责人: GEORGE M. CARMAN
• 金额: $ 7.65万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-03 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7889114
• 关键词: Address; Affect; Affinity; Biochemical; Biological; CDPdiacylglycerol-inositol 3-phosphatidyltransferase; Cell membrane; Choline Kinase; ETV3 gene; Enzyme Gene; Enzymes; Ethanolamine kinase; Eukaryotic Cell; Genes; Genetic; Grant; Growth; Homeostasis; Mediating; Membrane; Metabolism; Mutation; Nutrient; Outcome; Pathway interactions; Phosphatidate Phosphatase; Phosphatidylserine Synthase; Phospholipid Metabolism; Phospholipids; Phosphoric Monoester Hydrolases; Physiological; Play; Property; Regulation; Repression; Research; Role; Saccharomyces cerevisiae; Stress; Triglycerides; Vesicle; Work; Yeasts; Zinc; deprivation; enzyme activity; enzyme pathway; mutant; phosphatidate; positional cloning; promoter; reconstitution; response; zinc-binding protein

The yeast Saccharomyces cerevisiae responds to a variety of stress conditions (e.g., zinc depletion) by regulating the expression of several enzyme activities including those involved in phospholipid synthesis. Zinc is an essential nutrient required for the growth and metabolism of S. cerevisiae, and of higher eukaryotic cells. The work proposed in this application will address mechanisms by which the expression of key Dhospholipid biosynthetic enzyme activities is regulated in response to zinc depletion. We will examine the lypothesis that zinc depletion induces a cytosolic Mg2+-dependent and NEM-sensitive phosphatidate phosphatase that reduces phosphatidate concentration in the ER. This in turn triggers the Opilp-mediated repression of CHO1 (encodes phosphatidylserine synthase) and other UAS|No-containing genes. This question will be addressed using a phosphatase mutant that will be isolated by a reverse genetic approach requiring the purification of the cytosolic Mg2+-dependent phosphatidate phosphatase enzyme. The regulation of expression of this phosphatase gene by zinc depletion will be examined, along with the biochemical properties of the purified enzyme. We will examine the hypothesis that E/C/7-encoded ethanolamine kinase and CK/t-encoded choline kinase are induced in response to zinc depletion. Mechanisms responsible for the zinc-mediated regulation of the EKI1 and CKI1 genes will be examined, and the biological relevance of regulation for each gene will be examined using selective promoter mutations that affect their expression. The biological relevance of the Opilp-mediated repression of CHO1 and the Zaplp- mediated induction of PIS1 (encodes phosphatidylinositol synthase) will be explored with mutants that are unable to down-regulate phosphatidylserine synthase or up-regulate phosphatidylinositol synthase activities, respectively, in response to zinc depletion. We will examine the hypothesis that the function of the high- affinity plasma membrane zinc transporter Zrt1 p is regulated by changes in phospholipid composition brought about by phospholipid synthesis regulation by zinc depletion. Zinc transport function will be addressed with mutants defective in phospholipid synthesis, and with Zrtlp reconstituted into unilamellar vesicles with varying phospholipid composition.

酿酒酵母通过以下方式响应各种应激条件（例如缺锌）： 调节多种酶活性的表达，包括那些参与磷脂合成的酶活性。 锌是酿酒酵母和高等真核生物生长和代谢所需的必需营养素 细胞。本申请中提出的工作将解决关键表达的机制 磷脂生物合成酶活性因锌消耗而受到调节。我们将检查 推测锌耗尽会诱导细胞质 Mg2+ 依赖性且 NEM 敏感的磷脂酸 降低内质网中磷脂酸盐浓度的磷酸酶。这反过来又触发了 Opilp 介导的 抑制 CHO1（编码磷脂酰丝氨酸合酶）和其他包含 UAS|No 的基因。这 将使用通过反向遗传方法分离的磷酸酶突变体来解决这个问题 需要纯化胞质 Mg2+ 依赖性磷脂酸磷酸酶。这 将检查锌消耗对该磷酸酶基因表达的调节，以及 纯化酶的生化特性。我们将检验 E/C/7 编码的假设 乙醇胺激酶和 CK/t 编码的胆碱激酶因锌耗尽而被诱导。 将检查负责锌介导的 EKI1 和 CKI1 基因调节的机制，以及 将使用选择性启动子突变来检查每个基因调控的生物学相关性， 影响他们的表达。 Opilp 介导的 CHO1 和 Zaplp- 抑制的生物学相关性 将用突变体探索 PIS1（编码磷脂酰肌醇合酶）介导的诱导 无法下调磷脂酰丝氨酸合酶或上调磷脂酰肌醇合酶活性， 分别响应锌耗尽。我们将检验这样的假设：高的功能 亲和质膜锌转运蛋白 Zrt1 p 通过磷脂组成的变化进行调节 由锌消耗调节磷脂合成引起。锌的转运功能 解决了磷脂合成缺陷的突变体，并用 Zrtlp 重构为单层 具有不同磷脂成分的囊泡。

项目成果

N-cadherin mediates the migration of MCF-10A cells undergoing bone morphogenetic protein 4-mediated epithelial mesenchymal transition.

N-钙粘蛋白介导 MCF-10A 细胞的迁移，经历骨形态发生蛋白 4 介导的上皮间质转化。

• DOI: 10.1007/s13277-014-2991-9
• 发表时间: 2015
• 期刊: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine
• 作者: Park,Ki-Sook;Dubon,MariaJose;Gumbiner,BarryM
• 通讯作者: Gumbiner,BarryM

CDPdiacylglycerol synthase from yeast.

来自酵母的 CDP 二酰基甘油合酶。

• DOI: 10.1016/0076-6879(92)09030-7
• 发表时间: 1992
• 期刊: Methods in enzymology
• 作者: Carman,GM;Kelley,MJ
• 通讯作者: Kelley,MJ

Regulation of phosphatidate phosphatase activity from the yeast Saccharomyces cerevisiae by sphingoid bases.

通过鞘氨醇碱调节酿酒酵母的磷脂酸磷酸酶活性。

• 发表时间: 1993
• 期刊: The Journal of biological chemistry
• 作者: Wu,WI;Lin,YP;Wang,E;MerrillJr,AH;Carman,GM
• 通讯作者: Carman,GM

Enzymes of phosphoinositide synthesis in secretory vesicles destined for the plasma membrane in Saccharomyces cerevisiae.

酿酒酵母中用于质膜的分泌囊泡中磷酸肌醇合成的酶。

• DOI: 10.1128/jb.172.7.4115-4117.1990
• 发表时间: 1990
• 期刊: Journal of bacteriology
• 影响因子: 3.2
• 作者: Kinney,AJ;Carman,GM
• 通讯作者: Carman,GM

Phospholipids chiral at phosphorus. Steric course of the reactions catalyzed by phosphatidylserine synthase from Escherichia coli and yeast.

• DOI: 10.1021/bi00387a042
• 发表时间: 1987-06
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: C. Raetz;G. Carman;W. Dowhan;R. Jiang;W. Waszkuć;W. Loffredo;M. Tsai