RudLOV是一种光学同步的货物运输方式,显示出意想不到的王朝效应。

IF 6.5 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY EMBO Reports Pub Date : 2024-12-10 DOI:10.1038/s44319-024-00342-z
Tatsuya Tago, Takumi Ogawa, Yumi Goto, Kiminori Toyooka, Takuro Tojima, Akihiko Nakano, Takunori Satoh, Akiko K Satoh
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引用次数: 0

摘要

分泌物的实时成像是了解膜运输机制的有力方法。诱导货物从细胞器同步释放是增强显微观察的关键。我们开发了一种光学货物释放方法,“使用LOV2暗状态保留”(RudLOV),可以在货物释放过程中精确控制空间,时间和数量。使用RudLOV的有限数量的货物释放能够可视化高尔基/跨高尔基网络上的货物储罐运动和货物特定出口地点。此外,通过使用RudLOV控制货物释放的时间,我们揭示了众所周知的动力蛋白抑制剂王朝的典型和非典型效应,它抑制早期高尔基转运,而不是晚期高尔基转运,并退出动力蛋白-2活跃的反式高尔基网络。在经过王朝处理的细胞中,高尔基体堆积顺侧的COPI囊泡的积累表明,王朝针对的是早期高尔基池或内质网的COPI脱包/系结/融合机制,而不是晚期高尔基池。这些结果为高尔基堆的蓄水池成熟提供了洞见。
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RudLOV is an optically synchronized cargo transport method revealing unexpected effects of dynasore.

Live imaging of secretory cargoes is a powerful method for understanding the mechanisms of membrane trafficking. Inducing the synchronous release of cargoes from an organelle is key for enhancing microscopic observation. We developed an optical cargo-releasing method, 'retention using dark state of LOV2' (RudLOV), which enables precise spatial, temporal, and quantity control during cargo release. A limited amount of cargo-release using RudLOV is able to visualize cargo cisternal-movement and cargo-specific exit sites on the Golgi/trans-Golgi network. Moreover, by controlling the timing of cargo-release using RudLOV, we reveal the canonical and non-canonical effects of the well-known dynamin inhibitor dynasore, which inhibits early- but not late-Golgi transport and exits from the trans-Golgi network where dynamin-2 is active. Accumulation of COPI vesicles at the cis-side of the Golgi stacks in dynasore-treated cells suggests that dynasore targets COPI-uncoating/tethering/fusion machinery in the early-Golgi cisternae or endoplasmic reticulum but not in the late-Golgi cisternae. These results provide insight into the cisternal maturation of Golgi stacks.

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来源期刊
EMBO Reports
EMBO Reports 生物-生化与分子生物学
CiteScore
11.20
自引率
1.30%
发文量
267
审稿时长
1 months
期刊介绍: EMBO Reports is a scientific journal that specializes in publishing research articles in the fields of molecular biology, cell biology, and developmental biology. The journal is known for its commitment to publishing high-quality, impactful research that provides novel physiological and functional insights. These insights are expected to be supported by robust evidence, with independent lines of inquiry validating the findings. The journal's scope includes both long and short-format papers, catering to different types of research contributions. It values studies that: Communicate major findings: Articles that report significant discoveries or advancements in the understanding of biological processes at the molecular, cellular, and developmental levels. Confirm important findings: Research that validates or supports existing knowledge in the field, reinforcing the reliability of previous studies. Refute prominent claims: Studies that challenge or disprove widely accepted ideas or hypotheses in the biosciences, contributing to the correction and evolution of scientific understanding. Present null data: Papers that report negative results or findings that do not support a particular hypothesis, which are crucial for the scientific process as they help to refine or redirect research efforts. EMBO Reports is dedicated to maintaining high standards of scientific rigor and integrity, ensuring that the research it publishes contributes meaningfully to the advancement of knowledge in the life sciences. By covering a broad spectrum of topics and encouraging the publication of both positive and negative results, the journal plays a vital role in promoting a comprehensive and balanced view of scientific inquiry. 
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