Wang Z, Wilson W A, Fujino M A, Roach P J
Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, 635 Barnhill Drive, Indianapolis, IN 46202, USA.
Mol Cell Biol. 2001 Sep;21(17):5742-52. doi: 10.1128/MCB.21.17.5742-5752.2001.
In the yeast Saccharomyces cerevisiae, glycogen is accumulated as a carbohydrate reserve when cells are deprived of nutrients. Yeast mutated in SNF1, a gene encoding a protein kinase required for glucose derepression, has diminished glycogen accumulation and concomitant inactivation of glycogen synthase. Restoration of synthesis in an snf1 strain results only in transient glycogen accumulation, implying the existence of other SNF1-dependent controls of glycogen storage. A genetic screen revealed that two genes involved in autophagy, APG1 and APG13, may be regulated by SNF1. Increased autophagic activity was observed in wild-type cells entering the stationary phase, but this induction was impaired in an snf1 strain. Mutants defective for autophagy were able to synthesize glycogen upon approaching the stationary phase, but were unable to maintain their glycogen stores, because subsequent synthesis was impaired and degradation by phosphorylase, Gph1p, was enhanced. Thus, deletion of GPH1 partially reversed the loss of glycogen accumulation in autophagy mutants. Loss of the vacuolar glucosidase, SGA1, also protected glycogen stores, but only very late in the stationary phase. Gph1p and Sga1p may therefore degrade physically distinct pools of glycogen. Pho85p is a cyclin-dependent protein kinase that antagonizes SNF1 control of glycogen synthesis. Induction of autophagy in pho85 mutants entering the stationary phase was exaggerated compared to the level in wild-type cells, but was blocked in apg1 pho85 mutants. We propose that Snf1p and Pho85p are, respectively, positive and negative regulators of autophagy, probably via Apg1 and/or Apg13. Defective glycogen storage in snf1 cells can be attributed to both defective synthesis upon entry into stationary phase and impaired maintenance of glycogen levels caused by the lack of autophagy.
在酿酒酵母中,当细胞缺乏营养时,糖原作为碳水化合物储备积累。SNF1基因发生突变的酵母,该基因编码葡萄糖去阻遏所需的蛋白激酶,其糖原积累减少且糖原合酶随之失活。在snf1菌株中恢复合成仅导致短暂的糖原积累,这意味着存在其他依赖SNF1的糖原储存控制机制。一项遗传筛选表明,参与自噬的两个基因APG1和APG13可能受SNF1调控。在进入稳定期的野生型细胞中观察到自噬活性增加,但在snf1菌株中这种诱导作用受损。自噬缺陷的突变体在接近稳定期时能够合成糖原,但无法维持其糖原储备,因为随后的合成受损且磷酸化酶Gph1p的降解增强。因此,缺失GPH1部分逆转了自噬突变体中糖原积累的损失。液泡葡萄糖苷酶SGA1的缺失也能保护糖原储备,但仅在稳定期后期起作用。因此,Gph1p和Sga1p可能降解物理上不同的糖原池。Pho85p是一种细胞周期蛋白依赖性蛋白激酶,它拮抗SNF1对糖原合成的控制。与野生型细胞相比,进入稳定期的pho85突变体中自噬的诱导作用更为明显,但在apg1 pho85突变体中受到阻断。我们提出,Snf1p和Pho85p分别是自噬的正调控因子和负调控因子,可能通过Apg1和/或Apg13起作用。snf1细胞中糖原储存缺陷可归因于进入稳定期时合成缺陷以及由于缺乏自噬导致的糖原水平维持受损。