Heintze Jacob, Costa Joana R, Weber Melanie, Ketteler Robin
MRC Laboratory for Molecular Cell Biology, University College London, London, UK.
MRC Laboratory for Molecular Cell Biology, University College London, London, UK.
Cell Signal. 2016 Sep;28(9):1380-1388. doi: 10.1016/j.cellsig.2016.06.015. Epub 2016 Jun 18.
Autophagy and cellular metabolism are tightly linked processes, but how individual metabolic enzymes regulate the process of autophagy is not well understood. This study implicates ribose-5-phosphate isomerase (RPIA), a key regulator of the pentose phosphate pathway, in the control of autophagy. We used a dual gene deletion strategy, combining shRNA-mediated knockdown studies with CRISPR/Cas9 genome editing. Knockdown of RPIA by shRNA or genomic deletion by CRISPR/Cas9 genome editing, results in an increase of ATG4B-mediated LC3 processing and in the appearance of LC3-positive autophagosomes in cells. Increased LC3 processing upon knockdown of RPIA can be reversed by treatment with the antioxidant N-acetyl cysteine. The results are consistent with a model in which RPIA suppresses autophagy and LC3 processing by modulation of redox signaling.
自噬与细胞代谢是紧密相连的过程，但个体代谢酶如何调节自噬过程尚不清楚。本研究表明，磷酸戊糖途径的关键调节因子5-磷酸核糖异构酶（RPIA）参与自噬的调控。我们采用了双基因缺失策略，将shRNA介导的敲低研究与CRISPR/Cas9基因组编辑相结合。通过shRNA敲低RPIA或通过CRISPR/Cas9基因组编辑进行基因缺失，会导致细胞中ATG4B介导的LC3加工增加以及LC3阳性自噬体的出现。用抗氧化剂N-乙酰半胱氨酸处理可逆转敲低RPIA后LC3加工增加的情况。这些结果与RPIA通过调节氧化还原信号抑制自噬和LC3加工的模型一致。
