Aberrant mitophagy has been implicated in a broad spectrum of disorders. PINK1, Parkin, and ubiquitin have pivotal roles in priming mitophagy. However, the entire regulatory landscape and the precise control mechanisms of mitophagy remain to be elucidated. Here, we uncover fundamental mitophagy regulation involving PINK1 and a non-canonical role of the mitochondrial Tu translation elongation factor (TUFm). The mitochondrion-cytosol dual-localized TUFm interacts with PINK1 biochemically and genetically, which is an evolutionarily conserved Parkin-independent route toward mitophagy. A PINK1-dependent TUFm phosphoswitch at Ser222 determines conversion from activating to suppressing mitophagy. PINK1 modulates differential translocation of TUFm because p-S222-TUFm is restricted predominantly to the cytosol, where it inhibits mitophagy by impeding Atg5-Atg12 formation. The self-antagonizing feature of PINK1/TUFm is critical for the robustness of mitophagy regulation, achieved by the unique kinetic parameters of p-S222-TUFm, p-S65-ubiquitin, and their common kinase PINK1. Our findings provide new mechanistic insights into mitophagy and mitophagy-associated disorders.

线粒体异常与多种疾病有关。PINK1，Parkin和泛素在引发线粒体吞噬中起关键作用。但是，尚需阐明线粒体的整个调控环境和精确的控制机制。在这里，我们发现涉及PINK1的基本线粒体调控和线粒体Tu翻译延伸因子（TUFm）的非典型作用。线粒体-胞质溶胶双定位的TUFm在生物化学和遗传上与PINK1相互作用，这是进化上保守的，独立于Parkin的线粒体途径。Ser222上依赖PINK1的TUFm磷酸开关决定了从激活到抑制线粒体的转化。PINK1调节TUFm的差异转运，因为p-S222-TUFm主要限于胞浆，它通过阻止Atg5-Atg12的形成来抑制线粒体。PINK1 / TUFm的自拮抗特征对于线粒体调控的鲁棒性至关重要，这是通过p-S222-TUFm，p-S65-泛素及其常见激酶PINK1的独特动力学参数实现的。我们的发现为线粒体和线粒体相关疾病提供了新的机制性见解。