Chang Yu-Yun, Neufeld Thomas P
Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455, USA.
Mol Biol Cell. 2009 Apr;20(7):2004-14. doi: 10.1091/mbc.e08-12-1250. Epub 2009 Feb 18.
The TOR kinases are conserved negative regulators of autophagy in response to nutrient conditions, but the signaling mechanisms are poorly understood. Here we describe a complex containing the protein kinase Atg1 and the phosphoprotein Atg13 that functions as a critical component of this regulation in Drosophila. We show that knockout of Atg1 or Atg13 results in a similar, selective defect in autophagy in response to TOR inactivation. Atg1 physically interacts with TOR and Atg13 in vivo, and both Atg1 and Atg13 are phosphorylated in a nutrient-, TOR- and Atg1 kinase-dependent manner. In contrast to yeast, phosphorylation of Atg13 is greatest under autophagic conditions and does not preclude Atg1-Atg13 association. Atg13 stimulates both the autophagic activity of Atg1 and its inhibition of cell growth and TOR signaling, in part by disrupting the normal trafficking of TOR. In contrast to the effects of normal Atg13 levels, increased expression of Atg13 inhibits autophagosome expansion and recruitment of Atg8/LC3, potentially by decreasing the stability of Atg1 and facilitating its inhibitory phosphorylation by TOR. Atg1-Atg13 complexes thus function at multiple levels to mediate and adjust nutrient-dependent autophagic signaling.
TOR激酶是响应营养状况的自噬保守负调控因子,但其信号传导机制尚不清楚。在这里,我们描述了一种包含蛋白激酶Atg1和磷蛋白Atg13的复合物,它是果蝇中这种调控的关键组成部分。我们表明,敲除Atg1或Atg13会导致在TOR失活时自噬出现类似的选择性缺陷。Atg1在体内与TOR和Atg13发生物理相互作用,并且Atg1和Atg13都以营养、TOR和Atg1激酶依赖性方式被磷酸化。与酵母不同,Atg13的磷酸化在自噬条件下最高,并且不排除Atg1-Atg13的结合。Atg13刺激Atg1的自噬活性及其对细胞生长和TOR信号传导的抑制,部分是通过破坏TOR的正常运输。与正常Atg13水平的作用相反,Atg13表达增加会抑制自噬体扩张和Atg8/LC3的募集,这可能是通过降低Atg1的稳定性并促进TOR对其进行抑制性磷酸化来实现的。因此,Atg1-Atg13复合物在多个水平上发挥作用,以介导和调节营养依赖性自噬信号传导。