Parkin drives pS65-Ub turnover independently of canonical autophagy in Drosophila.
Parkin drives pS65-Ub turnover independently of canonical autophagy in Drosophila.
复制标题
Parkin驱动果蝇中pS 65-Ub的周转独立于典型的自噬。
DOI：
10.15252/embr.202153552
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发表时间：
2022-12-06
期刊：
影响因子：
7.7
通讯作者：
中科院分区：
文献类型：
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作者：
帕金森病相关蛋白PINK 1和Parkin在维持线粒体质量控制的共同途径中起作用。虽然PINK 1-Parkin途径可以促进细胞培养中线粒体变性后的自噬线粒体周转（mitophagy），但提出了替代的质量控制途径。为了分析PINK 1-Parkin通路在体内的运作机制，我们开发了检测果蝇中Ser 65-磷酸化泛素（pS 65-Ub）的方法。暴露于氧化剂百草枯导致稳定的Pink 1依赖性pS 65-Ub产生，而pS 65-Ub在未受刺激的parkin-null果蝇中积累，与阻断的降解一致。此外，我们表明pS 65-Ub在体内特异性地积累在破坏的线粒体上。核心自噬蛋白Atg 1、Atg 5和Atg 8a的耗尽不会导致pS 65-Ub的积累达到与parkin缺失相同的程度，并且parkin的过表达促进了Atg 5缺失背景下基础和百草枯诱导的pS 65-Ub的转换。因此，我们已经确定pS 65-Ub免疫检测可用于分析体内Pink 1-Parkin功能，作为报告基因构建体的替代方案。此外，我们的研究结果表明，Pink 1-Parkin通路可以促进线粒体周转独立于体内的典型自噬。Pink 1介导的线粒体磷酸化泛素化（pS 65-Ub）在体内被氧化应激激活，选择性地在破坏的线粒体上积累，并通过parkin的作用独立于典型的自噬基因而降解。
Parkinson's disease‐related proteins, PINK1 and Parkin, act in a common pathway to maintain mitochondrial quality control. While the PINK1‐Parkin pathway can promote autophagic mitochondrial turnover (mitophagy) following mitochondrial toxification in cell culture, alternative quality control pathways are suggested. To analyse the mechanisms by which the PINK1–Parkin pathway operates in vivo, we developed methods to detect Ser65‐phosphorylated ubiquitin (pS65‐Ub) in Drosophila. Exposure to the oxidant paraquat led to robust, Pink1‐dependent pS65‐Ub production, while pS65‐Ub accumulates in unstimulated parkin‐null flies, consistent with blocked degradation. Additionally, we show that pS65‐Ub specifically accumulates on disrupted mitochondria in vivo. Depletion of the core autophagy proteins Atg1, Atg5 and Atg8a did not cause pS65‐Ub accumulation to the same extent as loss of parkin, and overexpression of parkin promoted turnover of both basal and paraquat‐induced pS65‐Ub in an Atg5‐null background. Thus, we have established that pS65‐Ub immunodetection can be used to analyse Pink1‐Parkin function in vivo as an alternative to reporter constructs. Moreover, our findings suggest that the Pink1‐Parkin pathway can promote mitochondrial turnover independently of canonical autophagy in vivo. Pink1‐mediated mitochondrial phospho‐ubiquitination (pS65‐Ub) is activated by oxidative stress in vivo, selectively accumulates on disrupted mitochondria, and is degraded via the action of parkin independently of canonical autophagy genes.
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影响因子：
64.8
作者：
Gladkova C;Maslen SL;Skehel JM;Komander D
通讯作者：
Komander D
影响因子：
64.8
作者：
Gan ZY;Callegari S;Cobbold SA;Cotton TR;Mlodzianoski MJ;Schubert AF;Geoghegan ND;Rogers KL;Leis A;Dewson G;Glukhova A;Komander D
通讯作者：
Komander D
影响因子：
5.5
作者：
Kocaturk NM;Gozuacik D
通讯作者：
Gozuacik D
影响因子：
16
作者：
Heo JM;Ordureau A;Paulo JA;Rinehart J;Harper JW
通讯作者：
Harper JW
DOI：
10.1083/jcb.201402104
发表时间：
2014-04-28
期刊：
The Journal of cell biology
影响因子：
--
作者：
Kane LA;Lazarou M;Fogel AI;Li Y;Yamano K;Sarraf SA;Banerjee S;Youle RJ
通讯作者：
Youle RJ