Fast-Growing Saccharomyces cerevisiae Cells with a Constitutive Unfolded Protein Response and Their Potential for Lipidic Molecule Production
Fast-Growing Saccharomyces cerevisiae Cells with a Constitutive Unfolded Protein Response and Their Potential for Lipidic Molecule Production
复制标题
具有组成型未折叠蛋白反应的快速生长的酿酒酵母细胞及其生产脂质分子的潜力
DOI：
10.1128/aem.01083-22
复制
发表时间：
2022
影响因子：
4.4
通讯作者：
Kimata Yukio
中科院分区：
文献类型：
--
作者：
Nguyen Phuong Thi Mai;Ishiwata-Kimata Yuki;Kimata Yukio
在酿酒酵母细胞中，内质网（ER）的功能障碍，即所谓的ER应激，导致HAC1mRNA转化为剪接形式（HAC1i），后者被翻译成转录因子，极大地改变了基因表达谱。这种细胞反应最终增强ER功能，并被称为未折叠蛋白反应（UPR）。因此，预期UPR的人工唤起将增加ER上和ER中的有益材料的生产率。然而，正如这里所证明的，细胞constitutively expressingHAC1i mRNA（HAC1i细胞），即使在非应激条件下也表现出很强的UPR，生长相当缓慢，经常产生快速生长和低UPR后代。有趣的是，在低浓度的内质网应激物衣霉素或细胞内表达位于内质网的绿色荧光蛋白（GFP）诱导的弱内质网应激存在下，HAC1i细胞生长更快，产生稳定的位于内质网的GFP的HAC1i细胞表现出强的UPR活性，携带高度扩增的内质网，并大量产生甘油三酯和异源类胡萝卜素。因此，我们建议，我们的研究结果提供了一个基础，代谢工程，以产生细胞产生有价值的peptidic molecules.IMPORTANCEThe UPR被认为是一种细胞反应，以科普未折叠的蛋白质在ER中的积累。In S.在酿酒酵母细胞中，UPR在非应激条件下被严重抑制。这项研究的结果阐明了UPR的严格调节的生理意义。组成型UPR诱导引起相当大的生长迟缓，这是部分获救的诱导弱ER应力。因此，我们推测，当UPR在缺乏异常ER客户蛋白的非应激细胞中被不适当地诱导时，UPR不适当地损害正常的细胞功能。本研究的另一个重点是S.酿酒酵母菌株稳定地表现出强UPR活性并大量产生甘油三酯和异源类胡萝卜素。我们预计我们的发现可能会应用于使用酵母细胞作为潜在的下一代技术来生产有价值的脂质分子。
In Saccharomyces cerevisiae cells, dysfunction of the endoplasmic reticulum (ER), so-called ER stress, leads to conversion ofHAC1mRNA to the spliced form (HAC1i), which is translated into a transcription factor that drastically changes the gene expression profile. This cellular response ultimately enhances ER functions and is named the unfolded protein response (UPR). Artificial evocation of the UPR is therefore anticipated to increase productivity of beneficial materials on and in the ER. However, as demonstrated here, cells constitutively expressingHAC1i mRNA (HAC1i cells), which exhibited a strong UPR even under nonstress conditions, grew considerably slowly and frequently yielded fast-growing and low-UPR progeny. Intriguingly, growth ofHAC1i cells was faster in the presence of weak ER stress that was induced by low concentrations of the ER stressor tunicamycin or by cellular expression of the ER-located version of green fluorescent protein (GFP).HAC1i cells producing ER-localized GFP stably exhibited a strong UPR activity, carried a highly expanded ER, and abundantly produced triglycerides and heterogenous carotenoids. We therefore propose that our findings provide a basis for metabolic engineering to generate cells producing valuable lipidic molecules.IMPORTANCEThe UPR is thought to be a cellular response to cope with the accumulation of unfolded proteins in the ER. In S. cerevisiae cells, the UPR is severely repressed under nonstress conditions. The findings of this study shed light on the physiological significance of the tight regulation of the UPR. Constitutive UPR induction caused considerable growth retardation, which was partly rescued by the induction of weak ER stress. Therefore, we speculate that when the UPR is inappropriately induced in unstressed cells lacking aberrant ER client proteins, the UPR improperly impairs normal cellular functions. Another important point of this study was the generation of S. cerevisiae strains stably exhibiting a strong UPR activity and abundantly producing triglycerides and heterogenous carotenoids. We anticipate that our findings may be applied to produce valuable lipidic molecules using yeast cells as a potential next-generation technique.