System biologic approach to lipid traffic and conversion in eukaryotic microorganisms and its application

真核微生物脂质运输和转化的系统生物学方法及其应用

基本信息

批准号：
17208008
负责人：
OHTA Akinori
金额：
$ 31.37万
依托单位：
The university of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2007
项目状态：
已结题

项目摘要

This project was comprised of two parts : One is the analysis on the uptake and acyl-chain remodeling of phospholipids with short fatty acid residues by yeast Saccharomyces cerevisiae. In previous research, we showed that phosphatidylethanolamine (PE) and phosphatidylcholine (PC) with 8-to 12-carbon-fatty acid residues were incorporated and utilized by S. cerevisiae mutants defective in the synthesis of these phospholipids. In this project, we proved that these soluble phospholipids were. transported into the cells and their acyl chains are actually replaced with those of normal length by tracing the change of stable isotope-labeled PE and PC with decanoic acids. Thus the remodeling of phospholipids in their fatty acid residues is a common property of eukaryotic cells.The other part is the finding of a negative transcription factor, Yas3, for alkane-induced gene expression in Yarrowia lipolytica. In our previous research, we reported two cooperative positive regulatory factors, Yaslp and Yas2p, with bHLH motifs. We found that Yas3p changed its intracellular localization. When the lipolytica cells were grown in the presence of alkane, Yas3p-GFP was found in ER, but in the absence of alkane, it was found in nucleus, where it likely represses the function of Yas1p and Yas2p by binding to Yas2p. We also found Yas3p specifically bound to phosphatidic acid and PA phosphatase mutation increased expression of the alkane-inducible ALK1 gene, suggesting the change in the amount of PA is important in the function of Yas3p. We also investigated expression of 12 ALK genes by RT-PCR and found not all ALK genes are responsive to alkane.The latter part of our project resulted in a discovery of a very strong inducible promoter in Y. lipolytica and a mutant strain that is suitable for the expression of cytochrome P450 genes.

该项目由两个部分组成：一个是对摄入量和抗磷脂的分析，并重塑了用短脂肪酸残基对酵母菌酿酒酵母的磷脂的分析。在先前的研究中，我们表明，磷脂酰乙醇胺（PE）和磷脂酰胆碱（PC）具有8至12-碳脂肪酸残基，并被酿酒酵母突变体有缺陷并在这些磷脂的合成中使用。在这个项目中，我们证明了这些可溶性磷脂。通过追踪稳定的同位素标记的PE和PC的变化，用十烷基酸来取代进入细胞，其酰基链实际上被正常长度取代。因此，其脂肪酸残基中磷脂的重塑是真核细胞的常见特性。另一部分是脂溶液中烷烃诱导的基因表达的阴性转录因子YAS3的发现。在我们先前的研究中，我们报告了两个合作的阳性调节因素，YASLP和YAS2P，具有BHLH图案。我们发现YAS3P改变了其细胞内定位。当在烷烃存在下生长脂溶细胞时，在ER中发现了YAS3P-GFP，但是在没有烷烃的情况下，它在核中发现，它很可能通过与YAS2P结合而抑制Yas1p和Yas2p的功能。我们还发现YAS3P专门与磷脂酸和PA磷酸酶突变结合增加了烷烃诱导的ALK1基因的表达，这表明PA量的变化在YAS3P的功能中很重要。我们还通过RT-PCR研究了12种ALK基因的表达，并发现并非所有ALK基因都对烷烃有反应。我们项目的后半部分导致在Y. y. y. y. lipolytica中发现了非常强大的诱导启动子，而突变型菌株适合于细胞色素P450基因的表达。

项目成果

期刊论文数量（39）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The Bax Inhibitor-1 needs a functional electron transport chain for cell death suppression

Bax Inhibitor-1 需要功能性电子传递链来抑制细胞死亡

DOI：
发表时间：
2007
期刊：
FEBS Lett. 581
影响因子：
0
作者：
R. Oshima;K. Yoshinaga;他5名
通讯作者：
他5名

Crystallization and preliminary X-ray analsis of CTP : phosphoethanolamine cytidylyltransferase(ECT)from Saccharomyces cerevisiae

酿酒酵母磷酸乙醇胺胞苷酰转移酶 (ECT) 的结晶和初步 X 射线分析

DOI：
发表时间：
2006
期刊：
Acta Cryst. F62
影响因子：
0
作者：
Jun Ohtsuka;Koji Nagata;Woo Ceol Lee;Yusuke Ono;Ryouichi Fukuda;Akinori Ohta;Masaru Tanokura
通讯作者：
Masaru Tanokura

Crystallization and preliminary X-ray analsis of CTP : phosphoethanolamine cytidylyltransferase(ECT)from Saccharomyces cerevisiae.

CTP 的结晶和初步 X 射线分析：来自酿酒酵母的磷酸乙醇胺胞苷酰转移酶（ECT）。