Optogenetic tools for inducing organelle membrane rupture.

IF 4 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Journal of Biological Chemistry Pub Date : 2025-04-01 Epub Date: 2025-03-18 DOI:10.1016/j.jbc.2025.108421
Yuto Nagashima, Tomoya Eguchi, Ikuko Koyama-Honda, Noboru Mizushima
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引用次数: 0

Abstract

Disintegration of organelle membranes induces various cellular responses and has pathological consequences, including autoinflammatory diseases and neurodegeneration. Establishing methods to induce membrane rupture of specific organelles is essential to analyze the downstream effects of membrane rupture; however, the spatiotemporal induction of organelle membrane rupture remains challenging. Here, we develop a series of optogenetic tools to induce organelle membrane rupture by using engineered Bcl-2-associated X protein (BAX), which primarily functions to form membrane pores in the outer mitochondrial membrane (OMM) during apoptosis. When BAX is forced to target mitochondria, lysosomes, or the endoplasmic reticulum (ER) by replacing its C-terminal transmembrane domain (TMD) with organelle-targeting sequences, the BAX mutants rupture their targeted membranes. To regulate the activity of organelle-targeted BAX, the photosensitive light-oxygen-voltage-sensing 2 (LOV2) domain is fused to the N-terminus of BAX. The resulting LOV2-BAX fusion protein exhibits blue light-dependent membrane-rupture activity on various organelles, including mitochondria, the ER, and lysosomes. Thus, LOV2-BAX enables spatiotemporal induction of membrane rupture across a broad range of organelles, expanding research opportunities on the consequences of organelle membrane disruption.

诱导细胞器膜破裂的光遗传学工具。
细胞器膜的解体引起各种细胞反应,并具有病理后果,包括自身炎症性疾病和神经变性。建立诱导特定细胞器膜破裂的方法是分析膜破裂下游效应的必要条件;然而,细胞器膜破裂的时空诱导仍然具有挑战性。在这里,我们开发了一系列光遗传学工具,通过使用工程化的bcl -2相关X蛋白(BAX)诱导细胞器膜破裂,该蛋白主要作用是在细胞凋亡过程中在外线粒体膜(OMM)形成膜孔。当BAX通过细胞器靶向序列取代其c端跨膜结构域(TMD)而被迫靶向线粒体、溶酶体或内质网(ER)时,BAX突变体会破坏其靶向膜。为了调节细胞器靶向BAX的活性,光敏光氧电压传感2 (LOV2)结构域融合到BAX的n端。所得到的LOV2-BAX融合蛋白在各种细胞器上表现出蓝光依赖的膜破裂活性,包括线粒体、内质网和溶酶体。因此,LOV2-BAX能够在广泛的细胞器中时空诱导膜破裂,扩大了细胞器膜破坏后果的研究机会。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Biological Chemistry
Journal of Biological Chemistry Biochemistry, Genetics and Molecular Biology-Biochemistry
自引率
4.20%
发文量
1233
期刊介绍: The Journal of Biological Chemistry welcomes high-quality science that seeks to elucidate the molecular and cellular basis of biological processes. Papers published in JBC can therefore fall under the umbrellas of not only biological chemistry, chemical biology, or biochemistry, but also allied disciplines such as biophysics, systems biology, RNA biology, immunology, microbiology, neurobiology, epigenetics, computational biology, ’omics, and many more. The outcome of our focus on papers that contribute novel and important mechanistic insights, rather than on a particular topic area, is that JBC is truly a melting pot for scientists across disciplines. In addition, JBC welcomes papers that describe methods that will help scientists push their biochemical inquiries forward and resources that will be of use to the research community.
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