Role of the integral membrane protease ZMPSTE24 in membrane protein biogenesis and virus-host cell fusion

整合膜蛋白酶 ZMPSTE24 在膜蛋白生物发生和病毒-宿主细胞融合中的作用

• 批准号: 10622926
• 负责人: Susan D. Michaelis
• 金额: $ 51.09万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10622926
• 关键词: Antiviral Agents; Biogenesis; Biological; Biology; COVID-19 pandemic; Cell fusion; Cell membrane; Cells; Cellular Membrane; Cytoprotection; Defect; Disease; Endoplasmic Reticulum; Ensure; Environment; Goals; Health; Human; IFITM1 gene; Infection; Integral Membrane Protein; Interferons; Laboratories; Lamin Type A; Lead; Lipid Bilayers; Longevity; Longitudinal Studies; Membrane; Membrane Lipids; Membrane Proteins; Metalloproteases; Mus; Nuclear Matrix-Associated Proteins; Pathway interactions; Peptide Hydrolases; Positioning Attribute; Post-Translational Protein Processing; Premature aging syndrome; Progeria; Property; Proteins; Proteolytic Processing; Proteome; Quality Control; Research; Role; Viral; Viral Physiology; Virus; Virus Diseases; Zinc; antiviral drug development; biophysical properties; farnesylation; fluidity; insight; membrane activity; novel; overexpression; prelamin A; recruit; trafficking

PROJECT SUMMARY A significant fraction of the eukaryotic proteome is composed of integral membrane proteins, most of which are inserted and assembled at the endoplasmic reticulum (ER). The diverse biophysical characteristics, topology, posttranslational modifications, and activities of these membrane proteins necessitate distinct cellular pathways and numerous components to ensure their proper biogenesis and function. The goal of this project is to define a new mechanistic role of the zinc metalloprotease ZMPSTE24 in membrane protein biology. ZMPSTE24 has long been a research focus in my laboratory and is important for human health and longevity through its established role in the proteolytic processing of farnesylated prelamin A, precursor of the nuclear scaffold protein lamin A. Defects in this processing step lead to premature aging (progeria) diseases. However, an intriguing new function for ZMPSTE24 in viral defense was recently discovered by others and surprisingly does not require its catalytic activity: ZMPSTE24 confers potent antiviral activity against many enveloped viruses through its interaction with a class of small membrane proteins called interferon-induced transmembrane proteins (IFITM1, 2, and 3). The IFITMs block virus-host cell fusion by a mechanism that involves “rigidifying” host cell membranes. As is the case for IFITMs, the overexpression of ZMPSTE24 robustly protects cells from infection by enveloped viruses, and its proteolytic activity is not needed in this role. Furthermore, depletion of ZMPSTE24 in cells and mice cause them to succumb to viral infection. These findings place ZMPSTE24 at an important position in the cell’s first line of defense against viral infection, likely through a general cell biological role. Here we hypothesize that ZMPSTE24 defines a central component in a known or new pathway for membrane protein biogenesis (insertion, topology, stability/quality control, posttranslational modification, or oligomerization), with IFITM3 as its substrate. Alternatively, ZMPSTE24 may facilitate IFITM3’s membrane rigidifying function in some other way, directly by recruitment to IFITM3, or indirectly by altering the composition or properties of the lipid bilayer. This project represents an exciting new direction in my laboratory’s long-term studies of ZMPSTE24, inspired by the convergence of ZMPSTE24’s newly discovered role in viral defense and the COVID-19 pandemic. Nevertheless, the studies we propose also relate to earlier research in my laboratory on membrane protein topology, trafficking, and ER quality control. Deciphering the antiviral role of ZMPSTE24 via the IFITMs presents an intriguing puzzle that we are primed to solve. We expect the studies proposed here will uncover a new fundamental role(s) for ZMPSTE24 in membrane protein biogenesis or membrane lipid composition or fluidity. Furthermore, insight into the mechanism whereby ZMPSTE24 enables IFITMs to block virus-host-cell fusion could ultimately be harnessed to develop a novel anti-viral drug.

项目摘要 真核蛋白的很大一部分由整合性膜蛋白组成，其中大多数 在内质网（ER）插入并组装。潜水员生物物理特征， 这些膜蛋白的拓扑，翻译后修饰和活性必要的不同细胞 途径和众多成分，以确保其适当的生物发生和功能。这个项目的目标 是定义金属蛋白酶ZMPSTE24在膜蛋白中的新机械作用 生物学。 ZMPSTE24长期以来一直是我的实验室的研究重点，对人类健康和 通过其在Farnesyrated Prelamin A的蛋白水解加工中固定的作用，寿命 核脚手架蛋白层lamin A.在此加工步骤中的缺陷导致过早衰老（后代）疾病。 但是，其他人最近发现了ZMPSTE24在病毒防御中引人入胜的新功能， 出乎意料的是，不需要其催化活性：ZMPSTE24承认许多针对许多人的抗病毒活性 通过与一类称为干扰诱导的小膜蛋白的相互作用，包裹的病毒通过其相互作用 跨膜蛋白（IFITM1、2和3）。 IFITM通过一种机制阻止病毒宿主宿主融合 涉及“刚化”宿主细胞膜。与IFITM一样，ZMPSTE24的过表达 坚固地保护细胞免受包膜病毒的感染，并且在此作用中不需要其蛋白水解活性。 此外，细胞和小鼠中ZMPSTE24的耗竭导致它们屈服于病毒感染。这些 发现将ZMPSTE24置于电池对病毒感染的第一条防御线中的重要位置，可能 通过一般的细胞生物学作用。 在这里，我们假设ZMPSTE24定义了已知或新途径中的中心分量 膜蛋白生物发生（插入，拓扑，稳定/质量控制，翻译后 修改或寡聚化），以IFITM3为基材。或者，ZMPSTE24可能有助于 直接通过招募IFITM3或 通过改变脂质双层的组成或特性间接。这个项目代表了一个令人兴奋的 我实验室对ZMPSTE24的长期研究的新方向，灵感来自ZMPSTE24的融合 新发现的在病毒防御和COVID-19大流行中的作用。然而，我们提出的研究也 与我的实验室有关膜蛋白拓扑，贩运和ER质量控制的早期研究有关。 通过IFITMS删除ZMPSTE24的抗病毒作用，这是一个有趣的难题，我们已准备就绪 解决。我们希望这里提出的研究将发现ZMPSTE24在 膜蛋白生物发生或膜脂质组成或流动性。此外，洞察 ZMPSTE24使IFITM能够阻止病毒宿主融合的机制最终可以利用 开发一种新型的抗病毒药物。

项目成果

The farnesyl transferase inhibitor (FTI) lonafarnib improves nuclear morphology in ZMPSTE24-deficient fibroblasts from patients with the progeroid disorder MAD-B.

• DOI: 10.1080/19491034.2023.2288476
• 发表时间: 2023-12
• 期刊: Nucleus (Austin, Tex.)

Site specificity determinants for prelamin A cleavage by the zinc metalloprotease ZMPSTE24.

• DOI: 10.1074/jbc.ra120.015792
• 发表时间: 2021-01
• 期刊: The Journal of biological chemistry
• 作者: Babatz TD;Spear ED;Xu W;Sun OL;Nie L;Carpenter EP;Michaelis S
• 通讯作者: Michaelis S

Defining substrate requirements for cleavage of farnesylated prelamin A by the integral membrane zinc metalloprotease ZMPSTE24.

• DOI: 10.1371/journal.pone.0239269
• 发表时间: 2020
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Wood KM;Spear ED;Mossberg OW;Odinammadu KO;Xu W;Michaelis S
• 通讯作者: Michaelis S

Prelamin A and ZMPSTE24 in premature and physiological aging.

• DOI: 10.1080/19491034.2023.2270345
• 发表时间: 2023-12
• 期刊: Nucleus (Austin, Tex.)

Abolishing the prelamin A ZMPSTE24 cleavage site leads to progeroid phenotypes with near-normal longevity in mice.

• DOI: 10.1073/pnas.2118695119
• 发表时间: 2022-03-01
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Wang Y;Shilagardi K;Hsu T;Odinammadu KO;Maruyama T;Wu W;Lin CS;Damoci CB;Spear ED;Shin JY;Hsu W;Michaelis S;Worman HJ
• 通讯作者: Worman HJ