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Interorganellar Communication Through Membrane Contact Sites in Toxoplasma Gondii. 弓形虫通过膜接触点进行细胞间通信
Pub Date : 2023-08-06 eCollection Date: 2023-01-01 DOI: 10.1177/25152564231189064
Diego Huet, Silvia N J Moreno

Apicomplexan parasites are a group of protists that cause disease in humans and include pathogens like Plasmodium spp., the causative agent of malaria, and Toxoplasma gondii, the etiological agent of toxoplasmosis and one of the most ubiquitous human parasites in the world. Membrane contact sites (MCSs) are widespread structures within eukaryotic cells but their characterization in apicomplexan parasites is only in its very beginnings. Basic biological features of the T. gondii parasitic cycle support numerous organellar interactions, including the transfer of Ca2+ and metabolites between different compartments. In T. gondii, Ca2+ signals precede a series of interrelated molecular processes occurring in a coordinated manner that culminate in the stimulation of key steps of the parasite life cycle. Calcium transfer from the endoplasmic reticulum to other organelles via MCSs would explain the precision, speed, and efficiency that is needed during the lytic cycle of T. gondii. In this short review, we discuss the implications of these structures in cellular signaling, with an emphasis on their potential role in Ca2+ signaling.

表面复合寄生虫是一类能导致人类疾病的原生生物,包括疟疾病原体疟原虫和弓形虫等病原体,后者是弓形虫病的病原体,也是世界上最普遍的人类寄生虫之一。膜接触位点(MCSs)是真核细胞内广泛存在的结构,但其在 apicomplexan 寄生虫中的特性研究才刚刚起步。淋球菌寄生周期的基本生物学特征支持许多细胞器之间的相互作用,包括 Ca2+ 和代谢物在不同区室之间的转移。在淋球菌体内,Ca2+ 信号先于一系列相互关联的分子过程以协调的方式发生,最终刺激寄生虫生命周期的关键步骤。钙从内质网通过MCS转移到其他细胞器可以解释淋球菌溶解周期中所需的精确度、速度和效率。在这篇简短的综述中,我们将讨论这些结构在细胞信号传导中的意义,重点是它们在 Ca2+ 信号传导中的潜在作用。
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引用次数: 0
Bacterial AsmA-Like Proteins: Bridging the Gap in Intermembrane Phospholipid Transport. 细菌 AsmA 样蛋白:弥补膜间磷脂运输的差距
Pub Date : 2023-07-12 eCollection Date: 2023-01-01 DOI: 10.1177/25152564231185931
Sujeet Kumar, Natividad Ruiz

In eukaryotic cells, nonvesicular lipid transport between organelles is mediated by lipid-transfer proteins. Recently, a new class of these lipid transporters has been described to facilitate the bulk of inter-organelle lipid transport at contact sites by forming bridge-like structures with a hydrophobic groove through which lipids travel. Because their predicted structure is composed of repeating β-groove (RBG) domains, they have been named the RBG protein superfamily. Early studies on RBG proteins VPS13 and ATG2 recognized the resemblance of their predicted structures to that of the bacterial Lpt system, which transports newly synthesized lipopolysaccharides (LPS) between the inner and the outer membranes (IMs and OMs) of Gram-negative bacteria. In these didermic bacteria, the IMs and OMs are separated by an aqueous periplasmic compartment that is traversed by a bridge-like structure built with β-jelly roll domains from several Lpt proteins that provides a hydrophobic groove for LPS molecules to travel across the periplasm. Despite structural and functional similarities between RBG proteins and the Lpt system, the bacterial AsmA-like protein family has recently emerged as the likely ancestor of RBG proteins and long sought-after transporters that facilitate the transfer of phospholipids from the IM to the OM. Here, we review our current understanding of the structure and function of bacterial AsmA-like proteins, mainly focusing on recent studies that have led to the proposal that AsmA-like proteins mediate the bulk of phospholipid transfer between the IMs and OMs.

在真核细胞中,细胞器之间的非囊泡脂质运输是由脂质转移蛋白介导的。最近,一类新的脂质转运蛋白被描述出来,它们通过形成具有疏水槽的桥状结构,使脂质通过疏水槽,从而促进了大部分细胞器间脂质在接触部位的转运。由于它们的预测结构由重复的 β 沟(RBG)结构域组成,因此被命名为 RBG 蛋白超家族。对 RBG 蛋白 VPS13 和 ATG2 的早期研究发现,它们的预测结构与细菌的 Lpt 系统相似,后者在革兰氏阴性细菌的内膜和外膜(IMs 和 OMs)之间运输新合成的脂多糖(LPS)。在这些真菌中,内膜和外膜被一个含水的周质区隔开,周质区由一个桥状结构穿过,该结构由多个 Lpt 蛋白的β-果冻卷结构域构建而成,为 LPS 分子穿过周质提供了一个疏水槽。尽管 RBG 蛋白和 Lpt 系统在结构和功能上有相似之处,但最近出现的细菌 AsmA 样蛋白家族可能是 RBG 蛋白的祖先,也是人们长期寻找的促进磷脂从 IM 转移到 OM 的转运体。在此,我们回顾了目前我们对细菌 AsmA 样蛋白的结构和功能的理解,主要侧重于最近的研究,这些研究提出 AsmA 样蛋白介导了大部分磷脂在 IM 和 OM 之间的转移。
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引用次数: 0
Bridge-Like Lipid Transfer Proteins (BLTPs) in C. elegans: From Genetics to Structures and Functions. 秀丽隐杆线虫桥状脂传递蛋白(BLTPs):从遗传学到结构和功能。
Pub Date : 2023-07-12 eCollection Date: 2023-01-01 DOI: 10.1177/25152564231186489
Taruna Pandey, Jianxiu Zhang, Bingying Wang, Dengke K Ma

In eukaryotic cells, lipid transfer can occur at membrane contact sites (MCS) to facilitate the exchange of various lipids between two adjacent cellular organelle membranes. Lipid transfer proteins (LTPs), including shuttle LTP or bridge-like LTP (BLTP), transport lipids at MCS and are critical for diverse cellular processes, including lipid metabolism, membrane trafficking, and cell signaling. BLTPs (BLTP1-5, including the ATG2 and VPS13 family proteins) contain lipid-accommodating hydrophobic repeating β-groove (RBG) domains that allow the bulk transfer of lipids through MCS. Compared with vesicular lipid transfer and shuttle LTP, BLTPs have been only recently identified. Their functions and regulatory mechanisms are currently being unraveled in various model organisms and by diverse approaches. In this review, we summarize the genetics, structural features, and biological functions of BLTP in the genetically tractable model organism C. elegans. We discuss our recent studies and findings on C. elegans LPD-3, a prototypical megaprotein ortholog of BLTP1, with identified lipid transfer functions that are evolutionarily conserved in multicellular organisms and in human cells. We also highlight areas for future research of BLTP using C. elegans and complementary model systems and approaches. Given the emerging links of BLTP to several human diseases, including Parkinson's disease and Alkuraya-Kučinskas syndrome, discovering evolutionarily conserved roles of BLTPs and their mechanisms of regulation and action should contribute to new advances in basic cell biology and potential therapeutic development for related human disorders.

在真核细胞中,脂质转移可以发生在膜接触位点(MCS),以促进两个相邻细胞器膜之间的各种脂质交换。脂质转移蛋白(LTP),包括穿梭LTP或桥状LTP(BLTP),在MCS下转运脂质,对多种细胞过程至关重要,包括脂质代谢、膜运输和细胞信号传导。BLTP(BLTP1-5,包括ATG2和VPS13家族蛋白)包含脂质调节疏水性重复β-沟(RBG)结构域,该结构域允许脂质通过MCS大量转移。与囊泡脂质转移和穿梭LTP相比,BLTP是最近才被发现的。它们的功能和调节机制目前正在各种模式生物中通过不同的方法被揭示。在这篇综述中,我们总结了BLTP在遗传可处理的模式生物秀丽隐杆线虫中的遗传学、结构特征和生物学功能。我们讨论了我们最近对秀丽隐杆线虫LPD-3的研究和发现,LPD-3是BLTP1的原型巨蛋白同源物,具有在多细胞生物和人类细胞中进化保守的脂质转移功能。我们还强调了未来使用秀丽隐杆线虫和互补模型系统和方法研究BLTP的领域。鉴于BLTP与包括帕金森病和Alkuraya-Kučinskas综合征在内的几种人类疾病之间的新联系,发现BLTP在进化上的保守作用及其调节和作用机制将有助于基础细胞生物学的新进展和相关人类疾病的潜在治疗发展。
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引用次数: 0
IP3R at ER-Mitochondrial Contact Sites: Beyond the IP3R-GRP75-VDAC1 Ca2+ Funnel. ER-线粒体接触点的 IP3R:超越 IP3R-GRP75-VDAC1 Ca2+ 漏斗。
Pub Date : 2023-06-22 eCollection Date: 2023-01-01 DOI: 10.1177/25152564231181020
Peace Atakpa-Adaji, Adelina Ivanova

Membrane contact sites (MCS) circumvent the topological constraints of functional coupling between different membrane-bound organelles by providing a means of communication and exchange of materials. One of the most characterised contact sites in the cell is that between the endoplasmic reticulum and the mitochondrial (ERMCS) whose function is to couple cellular Ca2+ homeostasis and mitochondrial function. Inositol 1,4,5-trisphosphate receptors (IP3Rs) on the ER, glucose-regulated protein 75 (GRP 75) and voltage-dependent anion channel 1 (VDAC1) on the outer mitochondrial membrane are the canonical component of the Ca2+ transfer unit at ERMCS. These are often reported to form a Ca2+ funnel that fuels the mitochondrial low-affinity Ca2+ uptake system. We assess the available evidence on the IP3R subtype selectivity at the ERMCS and consider if IP3Rs have other roles at the ERMCS beyond providing Ca2+. Growing evidence suggests that all three IP3R subtypes can localise and regulate Ca2+ signalling at ERMCS. Furthermore, IP3Rs may be structurally important for assembly of the ERMCS in addition to their role in providing Ca2+ at these sites. Evidence that various binding partners regulate the assembly and Ca2+ transfer at ERMCS populated by IP3R-GRP75-VDAC1, suggesting that cells have evolved mechanisms that stabilise these junctions forming a Ca2+ microdomain that is required to fuel mitochondrial Ca2+ uptake.

膜接触点(MCS)为不同膜结合细胞器之间的功能耦合提供了沟通和物质交换的途径,从而规避了拓扑学上的限制。细胞中最具特征的接触点之一是内质网和线粒体之间的接触点(ERMCS),其功能是将细胞钙离子平衡和线粒体功能联系起来。ER 上的 1,4,5-三磷酸肌醇受体(IP3Rs)、线粒体外膜上的葡萄糖调节蛋白 75(GRP 75)和电压依赖性阴离子通道 1(VDAC1)是 ERMCS 上 Ca2+ 转移单元的典型组成部分。据报道,它们通常形成一个 Ca2+ 漏斗,为线粒体低亲和性 Ca2+ 摄取系统提供动力。我们评估了有关 ERMCS 上 IP3R 亚型选择性的现有证据,并考虑了 IP3R 在 ERMCS 上除了提供 Ca2+ 之外是否还有其他作用。越来越多的证据表明,所有三种 IP3R 亚型都能在 ERMCS 定位并调节 Ca2+ 信号。此外,IP3R 除了在 ERMCS 上提供 Ca2+ 外,在结构上可能对 ERMCS 的组装也很重要。有证据表明,在由 IP3R-GRP75-VDAC1 构成的 ERMCS 上,各种结合伙伴可调节其组装和 Ca2+ 传输,这表明细胞已进化出稳定这些连接的机制,从而形成一个 Ca2+ 微域,这是促进线粒体 Ca2+ 摄取所必需的。
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引用次数: 0
How Membrane Contact Sites Shape the Phagophore. 膜接触位点如何塑造Phagophore。
Pub Date : 2023-05-09 eCollection Date: 2023-01-01 DOI: 10.1177/25152564231162495
Cristina Capitanio, Anna Bieber, Florian Wilfling

During macroautophagy, phagophores establish multiple membrane contact sites (MCSs) with other organelles that are pivotal for proper phagophore assembly and growth. In S. cerevisiae, phagophore contacts have been observed with the vacuole, the ER, and lipid droplets. In situ imaging studies have greatly advanced our understanding of the structure and function of these sites. Here, we discuss how in situ structural methods like cryo-CLEM can give unprecedented insights into MCSs, and how they help to elucidate the structural arrangements of MCSs within cells. We further summarize the current knowledge of the contact sites in autophagy, focusing on autophagosome biogenesis in the model organism S. cerevisiae.

在大自噬过程中,吞噬体与其他细胞器建立多个膜接触位点(MCS),这些细胞器对吞噬体的正确组装和生长至关重要。在酿酒酵母中,已经观察到吞噬体与液泡、内质网和脂滴的接触。原位成像研究极大地促进了我们对这些位点的结构和功能的理解。在这里,我们讨论了冷冻CLEM等原位结构方法如何对MCS提供前所未有的见解,以及它们如何有助于阐明MCS在细胞内的结构安排。我们进一步总结了目前关于自噬中接触位点的知识,重点是模式生物酿酒酵母中的自噬体生物发生。
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引用次数: 2
VPS13 Forum Proceedings: XK, XK-Related and VPS13 Proteins in Membrane Lipid Dynamics. VPS13 论坛论文集:膜脂动态中的 XK、XK 相关蛋白和 VPS13 蛋白。
Pub Date : 2023-02-23 eCollection Date: 2023-01-01 DOI: 10.1177/25152564231156994
Kevin Peikert, Adrian Danek

In 2020, the pandemic interrupted the series of biannual International Neuroacanthocytosis Meetings that brought together clinicians, scientists, and patient groups to share research into a small group of devastating genetic diseases that combine both acanthocytosis (deformed red blood cells) and neurodegeneration with movement disorders. This Meeting Report describes talks at the 5th VPS13 Forum in January 2022, one of a series of online meetings held to fill the gap. The meeting addressed the basic biology of two key proteins implicated in chorea-acanthocytosis (mutations in VPS13A) and McLeod syndrome (mutations in XK). In a remarkable confluence of ideas, the speakers described different aspects of a single functional unit that comprises of VPS13A and XK proteins working together. Conditions caused by VPS13 (A-D) gene family mutations and related genes, such as XK, previously footnote knowledge, seem to turn central for a novel disease paradigm: bulk lipid transfer disorders.

2020 年,大流行病中断了一年两次的国际神经棘细胞病系列会议,这些会议将临床医生、科学家和患者团体聚集在一起,分享对一小部分兼具棘细胞病(畸形红细胞)和神经变性与运动障碍的破坏性遗传疾病的研究成果。本会议报告介绍了 2022 年 1 月举行的第五届 VPS13 论坛的会谈情况,该论坛是为填补这一空白而举行的一系列在线会议之一。会议讨论了与舞蹈棘细胞症(VPS13A 基因突变)和麦克劳德综合征(XK 基因突变)有关的两个关键蛋白的基础生物学问题。演讲者从不同角度阐述了由 VPS13A 蛋白和 XK 蛋白共同作用的单一功能单元,可谓思想交汇。由 VPS13(A-D)家族基因突变和相关基因(如 XK)引起的病症似乎成为一种新型疾病范例的核心:大量脂质转移障碍。
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引用次数: 0
An Expansion of the Endoplasmic Reticulum that Halts Autophagy is Permissive to Genome Instability. 阻止自噬的内质网扩张会导致基因组不稳定
Pub Date : 2023-02-22 eCollection Date: 2023-01-01 DOI: 10.1177/25152564231157706
Eliana Lara-Barba, Alba Torán-Vilarrubias, María Moriel-Carretero

The links between autophagy and genome stability, and whether they are important for lifespan and health, are not fully understood. We undertook a study to explore this notion at the molecular level using Saccharomyces cerevisiae. On the one hand, we triggered autophagy using rapamycin, to which we exposed mutants defective in preserving genome integrity, then assessed their viability, their ability to induce autophagy and the link between these two parameters. On the other hand, we searched for molecules derived from plant extracts known to have powerful benefits on human health to try to rescue the negative effects rapamycin had against some of these mutants. We uncover that autophagy execution is lethal for mutants unable to repair DNA double strand breaks, while the extract from Silybum marianum seeds induces an expansion of the endoplasmic reticulum (ER) that blocks autophagy and protects them. Our data uncover a connection between genome integrity and homeostasis of the ER whereby ER stress-like scenarios render cells tolerant to sub-optimal genome integrity conditions.

自噬与基因组稳定性之间的联系,以及它们对寿命和健康是否重要,目前还不完全清楚。我们利用酿酒酵母开展了一项研究,在分子水平上探索这一概念。一方面,我们使用雷帕霉素诱导自噬,使基因组完整性缺陷的突变体暴露在雷帕霉素下,然后评估它们的活力、诱导自噬的能力以及这两个参数之间的联系。另一方面,我们从已知对人类健康有强大益处的植物提取物中寻找分子,试图挽救雷帕霉素对其中一些突变体的负面影响。我们发现,对于无法修复DNA双链断裂的突变体来说,自噬的执行是致命的,而水飞蓟种子的提取物能诱导内质网(ER)的扩张,从而阻止自噬并保护突变体。我们的数据揭示了基因组完整性与内质网平衡之间的联系,在这种情况下,类似内质网应激的情景会使细胞对次优基因组完整性条件产生耐受性。
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引用次数: 0
E-Syt1 Regulates Neuronal Activity-Dependent Endoplasmic Reticulum-Plasma Membrane Junctions and Surface Expression of AMPA Receptors. E-Syt1调节神经元活动依赖的内质网-质膜连接和AMPA受体的表面表达。
Pub Date : 2023-01-01 DOI: 10.1177/25152564231185011
Ranran Mao, Chunfang Tong, Jia-Jia Liu

Endoplasmic reticulum (ER)-plasma membrane (PM) contact sites/junctions play important roles in cell physiology including signal transduction, ion and lipid transfer, and membrane dynamics. However, little is known about the dynamic regulation and functional roles of ER-PM junctions in neurons. Using a split green fluorescent protein-based membrane contact probe, we find that the density of ER-PM contact sites changes dynamically in the dendrites of hippocampal neurons undergoing long-term synaptic potentiation (LTP). We show that the Ca2±-sensing membrane tethering protein Extended Synaptotagmin 1 (E-Syt1) mediates the formation of ER-PM contact sites during LTP. We also show that E-Syt1 is required for neuronal activity-dependent surface expression of the α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid-type glutamate receptors. These findings implicate ER-PM junctions in the regulation of neurotransmitter receptor trafficking and synaptic plasticity.

内质网(ER)-质膜(PM)接触点/连接点在细胞生理中起着重要的作用,包括信号转导、离子和脂质转移以及膜动力学。然而,对ER-PM连接在神经元中的动态调控和功能作用知之甚少。利用分离绿色荧光蛋白膜接触探针,我们发现在经历长期突触增强(LTP)的海马神经元树突中ER-PM接触位点的密度是动态变化的。我们发现Ca2±传感膜系泊蛋白扩展突触塔蛋白1 (E-Syt1)在LTP期间介导ER-PM接触位点的形成。我们还发现E-Syt1是神经元活性依赖的α-氨基-3-羟基-5-甲基-4-异恶唑丙酸型谷氨酸受体表面表达所必需的。这些发现暗示ER-PM连接在调节神经递质受体运输和突触可塑性。
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引用次数: 0
Complementation Assay Using Fusion of Split-GFP and TurboID (CsFiND) Enables Simultaneous Visualization and Proximity Labeling of Organelle Contact Sites in Yeast. 利用Split-GFP和TurboID (CsFiND)的融合进行互补分析,可以同时可视化和接近标记酵母细胞器接触位点。
Pub Date : 2023-01-01 DOI: 10.1177/25152564231153621
Shintaro Fujimoto, Shinya Tashiro, Yasushi Tamura

Numerous studies have revealed that organelle membrane contact sites (MCSs) play important roles in diverse cellular events, including the transport of lipids and ions between connected organelles. To understand MCS functions, it is essential to uncover proteins that accumulate at MCSs. Here, we develop a complementation assay system termed CsFiND (Complementation assay using Fusion of split-GFP and TurboID) for the simultaneous visualization of MCSs and identification of MCS-localized proteins. We express the CsFiND proteins on the endoplasmic reticulum and mitochondrial outer membrane in yeast to verify the reliability of CsFiND as a tool for identifying MCS-localized proteins.

大量研究表明,细胞器膜接触位点(MCSs)在多种细胞事件中发挥重要作用,包括脂质和离子在相连细胞器之间的运输。为了了解MCS的功能,有必要揭示MCS中积累的蛋白质。在这里,我们开发了一种名为CsFiND的互补分析系统(利用split-GFP和TurboID融合的互补分析),用于同时可视化mcs和鉴定mcs定位蛋白。我们在酵母内质网和线粒体外膜上表达了CsFiND蛋白,以验证CsFiND作为鉴定mcs定位蛋白的工具的可靠性。
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引用次数: 0
Evolutionary History of Oxysterol-Binding Proteins Reveals Complex History of Duplication and Loss in Animals and Fungi. 动物和真菌中氧化甾醇结合蛋白的进化史揭示了复制和丢失的复杂历史。
Pub Date : 2023-01-01 DOI: 10.1177/25152564221150428
Rohan P Singh, Yu-Ping Poh, Savar D Sinha, Jeremy G Wideman

Cells maintain the specific lipid composition of distinct organelles by vesicular transport as well as non-vesicular lipid trafficking via lipid transport proteins. Oxysterol-binding proteins (OSBPs) are a family of lipid transport proteins that transfer lipids at various membrane contact sites (MCSs). OSBPs have been extensively investigated in human and yeast cells where 12 have been identified in Homo sapiens and 7 in Saccharomyces cerevisiae. The evolutionary relationship between these well-characterized OSBPs is still unclear. By reconstructing phylogenies of eukaryote OSBPs, we show that the ancestral Saccharomycotina had four OSBPs, the ancestral fungus had five OSBPs, and the ancestral animal had six OSBPs, whereas the shared ancestor of animals and fungi as well as the ancestral eukaryote had only three OSBPs. Our analyses identified three undescribed ancient OSBP orthologues, one fungal OSBP (Osh8) lost in the lineage leading to yeast, one animal OSBP (ORP12) lost in the lineage leading to vertebrates, and one eukaryotic OSBP (OshEu) lost in both the animal and fungal lineages.

细胞通过囊泡转运以及通过脂质转运蛋白的非囊泡转运来维持不同细胞器的特定脂质组成。氧甾醇结合蛋白(osbp)是一个脂质转运蛋白家族,在不同的膜接触位点(MCSs)转移脂质。osbp已在人类和酵母细胞中进行了广泛的研究,其中在智人细胞中鉴定出12个,在酿酒酵母中鉴定出7个。这些特征明确的osbp之间的进化关系尚不清楚。通过重建真核生物osbp的系统发育,我们发现酵母菌的祖先有4个osbp,真菌的祖先有5个osbp,动物的祖先有6个osbp,而动物和真菌的共同祖先以及真核生物的祖先只有3个osbp。我们的分析确定了三个未描述的古代OSBP同源物,一个真菌OSBP (Osh8)在酵母谱系中丢失,一个动物OSBP (ORP12)在脊椎动物谱系中丢失,一个真核OSBP (OshEu)在动物和真菌谱系中都丢失。
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引用次数: 0
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