Dynamics of the Lipid Droplet Proteome of the Oleaginous Yeast Rhodosporidium toruloides
Dynamics of the Lipid Droplet Proteome of the Oleaginous Yeast Rhodosporidium toruloides
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DOI：
10.1128/ec.00141-14
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发表时间：
2015-01
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影响因子：
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通讯作者：
Zhiwei Zhu;Yunfeng Ding;Zhiwei Gong;Li Yang;Sufang Zhang;Congyan Zhang;Xinping Lin;Hongwei Shen-Hongwei-She
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文献类型：
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作者：
Zhiwei Zhu;Yunfeng Ding;Zhiwei Gong;Li Yang;Sufang Zhang;Congyan Zhang;Xinping Lin;Hongwei Shen-Hongwei-She
摘要 脂滴（LD）是普遍存在的细胞器，充当中性脂质储存库和脂质代谢的枢纽。操纵含油物种中 LD 的形成、进化和动员可能会导致脂肪酸衍生的生物燃料和化学品的生产。然而，调节 LD 动态的关键因素仍然知之甚少。在这里，我们从在营养丰富、氮限制和磷限制条件下培养的产脂酵母圆红冬孢酵母的细胞中纯化了 LD，并鉴定了 LD 相关蛋白。 LD蛋白质组由226个蛋白质组成，其中许多蛋白质参与脂质代谢和LD形成和进化。对我们之前比较转录组和蛋白质组数据集的进一步分析表明，85 个基因的转录水平和 77 个蛋白质的蛋白质丰度在营养限制条件下发生了变化。这种变化与脂质积累高度相关，并通过逆转录定量 PCR 得到部分证实。我们证明了主要的 LD 结构蛋白 Ldp1 是一种在富含脂质的细胞中上调的 LD 标记蛋白。当在酿酒酵母中过表达时，Ldp1 定位于 LD 表面并促进巨大 LD 的形成，表明 Ldp1 在控制 LD 动力学中发挥着重要作用。我们的结果显着促进了对产油酵母中脂质过量生产和储存的分子基础的理解，并且对于开发优质脂质生产者具有重要意义。
ABSTRACT Lipid droplets (LDs) are ubiquitous organelles that serve as a neutral lipid reservoir and a hub for lipid metabolism. Manipulating LD formation, evolution, and mobilization in oleaginous species may lead to the production of fatty acid-derived biofuels and chemicals. However, key factors regulating LD dynamics remain poorly characterized. Here we purified the LDs and identified LD-associated proteins from cells of the lipid-producing yeast Rhodosporidium toruloides cultured under nutrient-rich, nitrogen-limited, and phosphorus-limited conditions. The LD proteome consisted of 226 proteins, many of which are involved in lipid metabolism and LD formation and evolution. Further analysis of our previous comparative transcriptome and proteome data sets indicated that the transcription level of 85 genes and protein abundance of 77 proteins changed under nutrient-limited conditions. Such changes were highly relevant to lipid accumulation and partially confirmed by reverse transcription-quantitative PCR. We demonstrated that the major LD structure protein Ldp1 is an LD marker protein being upregulated in lipid-rich cells. When overexpressed in Saccharomyces cerevisiae, Ldp1 localized on the LD surface and facilitated giant LD formation, suggesting that Ldp1 plays an important role in controlling LD dynamics. Our results significantly advance the understanding of the molecular basis of lipid overproduction and storage in oleaginous yeasts and will be valuable for the development of superior lipid producers.