线粒体蛋白质稳态对于细胞功能至关重要。分子伴侣 HSP60 对于细胞功能至关重要，HSP60 表达失调与癌症和糖尿病有关。少数报道的携带 HSP60 基因变异的患者表现出神经发育迟缓和脑髓鞘形成不足。 Hsp60 与 260 多种线粒体蛋白相互作用，但受 HSP60 缺陷影响的线粒体蛋白和功能尚不清楚。我们研究了两种 HSP60 缺陷模型系统：(1) 携带可诱导显性失活 HSP60 突变蛋白的工程 HEK 细胞，(2) 斑马鱼HSP60 敲除幼虫。这两个系统均通过 RNASeq、蛋白质组学和靶向代谢组学以及与各自模型相关的几种功能测定进行了分析。此外，通过蛋白质组学分析了患有疾病相关 HSP60 变异的患者的皮肤成纤维细胞。我们发现，HSP60 缺乏会导致线粒体基质蛋白质组差异下调、应激反应转录激活以及胆固醇生物合成失调。这导致斑马鱼基因敲除幼虫体内脂质积累。我们的数据提供了 HSP60 缺陷对线粒体基质蛋白质组影响的概要。我们证明 HSP60 是线粒体功能和代谢途径的主要调节剂和调节剂。 HSP60 功能障碍还会影响细胞代谢并破坏综合应激反应。对胆固醇合成的影响解释了在携带 HSP60 遗传变异的患者中观察到的 HSP60 功能障碍对髓鞘形成的影响。版权所有 © 2024。由 Elsevier GmbH 出版。

Mitochondrial proteostasis is critical for cellular function. The molecular chaperone HSP60 is essential for cell function and dysregulation of HSP60 expression has been implicated in cancer and diabetes. The few reported patients carrying HSP60 gene variants show neurodevelopmental delay and brain hypomyelination. Hsp60 interacts with more than 260 mitochondrial proteins but the mitochondrial proteins and functions affected by HSP60 deficiency are poorly characterized.We studied two model systems for HSP60 deficiency: (1) engineered HEK cells carrying an inducible dominant negative HSP60 mutant protein, (2) zebrafish HSP60 knockout larvae. Both systems were analyzed by RNASeq, proteomics, and targeted metabolomics, and several functional assays relevant for the respective model. In addition, skin fibroblasts from patients with disease-associated HSP60 variants were analyzed by proteomics.We show that HSP60 deficiency leads to a differentially downregulated mitochondrial matrix proteome, transcriptional activation of stress responses, and dysregulated cholesterol biosynthesis. This leads to lipid accumulation in zebrafish knockout larvae.Our data provide a compendium of the effects of HSP60 deficiency on the mitochondrial matrix proteome. We show that HSP60 is a master regulator and modulator of mitochondrial functions and metabolic pathways. HSP60 dysfunction also affects cellular metabolism and disrupts the integrated stress response. The effect on cholesterol synthesis explains the effect of HSP60 dysfunction on myelination observed in patients carrying genetic variants of HSP60.Copyright © 2024. Published by Elsevier GmbH.