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.**

细胞器间的接触部位几乎存在于细胞的所有隔室之间，由于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，因此它们的研究已经成为非常热门的课题。细胞器间接触的一种最典型的类型是发生在线粒体和更平滑的细胞器之间的接触，它用于交换钙并调节呼吸、钙稳态和信号传递、磷脂生物合成等。然而，线粒体也可以被发现与含有核糖体的粗糙器相关，但这种联系的功能尚不清楚，这种线粒体相关粗糙器(mt-rer)是否是一种特殊类型的内质网仍有待确定。*在这里，我们分享大量令人信服的初步数据，以提出一个研究计划，目的是证明mt-rer是一种结构上不同的内质网类型，它通过翻译和分隔胆固醇和脂肪酸途径的酶和运输它们的脂蛋白来控制胆固醇和脂肪酸的合成。为了实现这个目标，我们将利用一种特别开发的方案，允许纯化小鼠肝脏mt-rer，并借鉴我们在低温电子显微镜、冷冻电子断层扫描、3D成像、转录组学和蛋白质组学表明：*1：mt-rer不连接到细胞的内质网*2：mt-rer有一个不同于ER的蛋白质组*3：光滑器和线粒体之间的接触部位(称为MAM或Merc)是mt-rer的一个亚域*4：脂质和胆固醇途径的蛋白质直接在mt-rer上翻译*5：脂类和胆固醇途径的蛋白质定位于mt-rer之间的界面和线粒体*6：线粒体包含一条活跃的胆固醇生物合成途径。这项研究将表明，线粒体和线粒体之间的并列作用是将脂质和胆固醇途径的底物、酶和前体分开，从而解释细胞如何有效地合成、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，因为他们将建立一种新颖的、结构和功能上不同的内质网类型；对于细胞生理学家，因为他们将赋予mt-rer通过合成脂蛋白使脂质和胆固醇在循环中的关键作用；以及对于MAM/MERC领域的研究人员，因为他们将提供一个框架来研究调节这个重要的亚细胞隔室的生物发生和重塑的机制。