Defects in autophagy lead to selective in vivo changes in turnover of cytosolic and organelle proteins in Arabidopsis.
Defects in autophagy lead to selective in vivo changes in turnover of cytosolic and organelle proteins in Arabidopsis.
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DOI：
10.1093/plcell/koac185
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发表时间：
2022-09-27
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自噬相关基因（ATG）突变体中自噬蛋白货物的鉴定由于蛋白质合成和蛋白质降解的变化而变得复杂。为了检测自噬货物，我们测量了拟南芥（Arabidopsis thaliana）atg5和atg11突变体的芽和根中的蛋白质降解率。这些数据表明，在丰度变化的蛋白质中，不到四分之一是可能的货物，并揭示了ATG11和ATG5在降解特定糖酵解酶和其他细胞质、叶绿体和ER驻留蛋白质中的作用，以及ATG11在降解线粒体和叶绿体蛋白质中的专门作用。在转化的原生质体中的蛋白定位和在抑制剂存在下的降解测定证实了自噬在降解糖酵解酶中的作用。自噬诱导磷酸盐（Pi）的限制改变代谢概况和蛋白质的合成和降解率的atg5和atg11植物。自噬突变体中氨基酸丰度的普遍降低和次级代谢产物的增加与特定蛋白质降解速率降低引起的catabolism改变和能量转换变化一致。结合蛋白质丰度和降解率的变化的措施，我们还确定了ATG11和ATG5相关的蛋白质货物的低PI诱导的自噬叶绿体和ER居民蛋白参与次生代谢。植物中自噬的蛋白质货物可以通过鉴定在自噬机制缺陷的突变体中丰度增加和降解速率降低的蛋白质来发现。
Identification of autophagic protein cargo in plants in autophagy-related genes (ATG) mutants is complicated by changes in protein synthesis and protein degradation. To detect autophagic cargo, we measured protein degradation rate in shoots and roots of Arabidopsis (Arabidopsis thaliana) atg5 and atg11 mutants. These data show that less than a quarter of proteins changing in abundance are probable cargo and revealed roles of ATG11 and ATG5 in degradation of specific glycolytic enzymes and of other cytosol, chloroplast, and ER-resident proteins, and a specialized role for ATG11 in degradation of proteins from mitochondria and chloroplasts. Protein localization in transformed protoplasts and degradation assays in the presence of inhibitors confirm a role for autophagy in degrading glycolytic enzymes. Autophagy induction by phosphate (Pi) limitation changed metabolic profiles and the protein synthesis and degradation rates of atg5 and atg11 plants. A general decrease in the abundance of amino acids and increase in secondary metabolites in autophagy mutants was consistent with altered catabolism and changes in energy conversion caused by reduced degradation rate of specific proteins. Combining measures of changes in protein abundance and degradation rates, we also identify ATG11 and ATG5-associated protein cargo of low Pi-induced autophagy in chloroplasts and ER-resident proteins involved in secondary metabolism. Protein cargo of autophagy in plants can be discovered by identifying proteins that increase in abundance and decrease in degradation rate in mutants deficient in autophagy machinery.
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