The mitochondrial rough-ER (mt-rER), a new type of endoplasmic reticulum: from structure to function

线粒体粗面-ER（mt-rER），一种新型内质网：从结构到功能

• 批准号: RGPIN-2017-06130
• 负责人: PELLEGRINI, Luca
• 金额: $ 2.04万
• 依托单位: Université Laval
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=661847
• 关键词: mitochondrial; rough; ER; mt; rER

Inter-organellar contact sites exist between nearly all the compartments of the cell, and their study has become very topical because more and more physiological and metabolic processes are coordinated by physical interactions between organelles. One of the best characterized type of inter-organellar contact is the one that occurs between mitochondria and the smoothER, which serves to exchange calcium and regulate respiration, Ca2+ homeostasis and signalling, phospholipid biosynthesis, and more. However, mitochondria can be found associated also to the ribosome-containing roughER, but the function of this association is unknown, and whether or not this mitochondria-associated roughER (mt-rER) is a specialized type of ER remains to be determined. ******Here, we share a wealth of compelling preliminary data to propose a research program that has the objective to show that the mt-rER is a structurally distinct type of ER that controls cholesterol and fatty acids synthesis by translating and compartmentalizing the enzymes of their pathway and the lipoproteins that traffic them.******To this goal, we will capitalize on an ad-hoc developed protocol allowing the purification of mouse liver mt-rER and on our experience in cryo-electron microscopy, cryo-electron tomography, 3D imaging, transcriptomics and proteomics to show that:******1: the mt-rER is not connected to the ER network of the cell ***2: the mt-rER has a proteome that is different from that of the ER***3: the contact site between the smoothER and the mitochondrion (known as MAM or MERC) is a subdomain of the mt-rER***4: proteins of the lipids and cholesterol pathway are translated directly at the mt-rER***5: proteins of the lipids and cholesterol pathway are localized at the interface between the mt-rER and the mitochondrion***6: the mt-rER contains an active cholesterol biosynthetic pathway.******Ultimately, this research will show that the juxtaposition between the mt-rER and the mitochondrion serves to compartmentalize substrates, enzymes and precursors of the lipid and cholesterol pathway, thereby explaining how the cell can efficiently synthesize, handle and transport huge amounts of these molecules.******These discoveries will have a major impact for many. For cell biologists, because they will establish the mt-rER as a novel, structurally and functionally distinct type of ER; for cell physiologists, because they will assigning to the mt-rER a key role in putting lipids and cholesterol in circulation through the synthesis of lipoproteins; and for researchers in the MAM/MERC field because they will provide a framework for studying the mechanisms regulating the biogenesis and remodelling of this important sub-cellular compartment.**

细胞器间的接触位点几乎存在于细胞的所有区室之间，并且他们的研究已变得非常热门，因为越来越多的生理和代谢过程是通过细胞器之间的物理相互作用来协调的。最典型的细胞器间接触类型之一是发生在线粒体和 smoothER 之间的接触，其作用是交换钙并调节呼吸、Ca2+ 稳态和信号传导、磷脂生物合成等。然而，线粒体也被发现与含有核糖体的roughER相关，但这种关联的功能尚不清楚，并且这种线粒体相关的rER（mt-rER）是否是一种特殊类型的ER仍有待确定。 ******在这里，我们分享大量令人信服的初步数据，以提出一项研究计划，其目标是证明 mt-rER 是一种结构独特的 ER 类型，通过翻译和划分其途径的酶以及运输它们的脂蛋白来控制胆固醇和脂肪酸的合成。********为了实现这一目标，我们将利用一项专门开发的协议，允许纯化小鼠肝脏 mt-rER 以及我们在冷冻电子显微镜、冷冻电子断层扫描、3D 成像、转录组学和蛋白质组学方面的经验表明：******1：mt-rER 未连接到细胞的 ER 网络***2：mt-rER 具有与 ER 不同的蛋白质组***3：smoothER 和线粒体（称为 MAM 或 MERC）之间的接触位点 是 mt-rER 的一个子域***4：脂质和胆固醇途径的蛋白质直接在 mt-rER 处翻译***5：脂质和胆固醇途径的蛋白质位于 mt-rER 和线粒体之间的界面***6：mt-rER 包含活性胆固醇生物合成途径。*******最终，本研究将 研究表明，mt-rER 和线粒体之间的并置可以区分脂质和胆固醇途径的底物、酶和前体，从而解释细胞如何有效合成、处理和运输大量这些分子。******这些发现将对许多人产生重大影响。对于细胞生物学家来说，因为他们将把 mt-rER 确立为一种新颖的、结构和功能独特的 ER 类型；对于细胞生理学家来说，因为他们认为 mt-rER 在通过脂蛋白的合成将脂质和胆固醇放入循环中发挥着关键作用；对于 MAM/MERC 领域的研究人员来说，因为它们将为研究调节这一重要亚细胞区室的生物发生和重塑的机制提供一个框架。**