Optogenetic cleavage of the Miro GTPase reveals the direct consequences of real-time loss of function in Drosophila.

IF 7.8 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY PLoS Biology Pub Date : 2023-08-17 eCollection Date: 2023-08-01 DOI:10.1371/journal.pbio.3002273
Francesca Mattedi, Ethlyn Lloyd-Morris, Frank Hirth, Alessio Vagnoni
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Abstract

Miro GTPases control mitochondrial morphology, calcium homeostasis, and regulate mitochondrial distribution by mediating their attachment to the kinesin and dynein motor complex. It is not clear, however, how Miro proteins spatially and temporally integrate their function as acute disruption of protein function has not been performed. To address this issue, we have developed an optogenetic loss of function "Split-Miro" allele for precise control of Miro-dependent mitochondrial functions in Drosophila. Rapid optogenetic cleavage of Split-Miro leads to a striking rearrangement of the mitochondrial network, which is mediated by mitochondrial interaction with the microtubules. Unexpectedly, this treatment did not impact the ability of mitochondria to buffer calcium or their association with the endoplasmic reticulum. While Split-Miro overexpression is sufficient to augment mitochondrial motility, sustained photocleavage shows that Split-Miro is surprisingly dispensable to maintain elevated mitochondrial processivity. In adult fly neurons in vivo, Split-Miro photocleavage affects both mitochondrial trafficking and neuronal activity. Furthermore, functional replacement of endogenous Miro with Split-Miro identifies its essential role in the regulation of locomotor activity in adult flies, demonstrating the feasibility of tuning animal behaviour by real-time loss of protein function.

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Miro-GTPase的光遗传学切割揭示了果蝇实时功能丧失的直接后果。
Miro GTP酶控制线粒体形态、钙稳态,并通过介导其与驱动蛋白和动力蛋白运动复合体的连接来调节线粒体分布。然而,目前尚不清楚Miro蛋白是如何在空间和时间上整合其功能的,因为尚未对蛋白功能进行急性破坏。为了解决这个问题,我们开发了一种光遗传学功能丧失的“分裂Miro”等位基因,用于精确控制果蝇的Miro依赖性线粒体功能。Split Miro的快速光遗传学切割导致线粒体网络的显著重排,这是由线粒体与微管的相互作用介导的。出乎意料的是,这种治疗并没有影响线粒体缓冲钙的能力或它们与内质网的结合。虽然Split-Miro过表达足以增强线粒体运动性,但持续的光切割表明,Split-Maro在维持线粒体加工能力方面是令人惊讶的可有可无。在体内成年果蝇神经元中,Split Miro光切割影响线粒体运输和神经元活动。此外,用Split Miro功能性取代内源性Miro确定了其在成年苍蝇运动活动调节中的重要作用,证明了通过蛋白质功能的实时丧失来调节动物行为的可行性。
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来源期刊
PLoS Biology
PLoS Biology 生物-生化与分子生物学
CiteScore
14.40
自引率
2.00%
发文量
359
审稿时长
3 months
期刊介绍: PLOS Biology is an open-access, peer-reviewed general biology journal published by PLOS, a nonprofit organization of scientists and physicians dedicated to making the world's scientific and medical literature freely accessible. The journal publishes new articles online weekly, with issues compiled and published monthly. ISSN Numbers: eISSN: 1545-7885 ISSN: 1544-9173
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