Reggiori Fulvio, Wang Chao-Wen, Stromhaug Per E, Shintani Takahiro, Klionsky Daniel J
Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109-1048, USA.
J Biol Chem. 2003 Feb 14;278(7):5009-20. doi: 10.1074/jbc.M210436200. Epub 2002 Nov 20.
Autophagy, pexophagy, and the Cvt pathway are processes that deliver hydrolytic enzymes and substrates to the yeast vacuole/lysosome via double-membrane cytosolic vesicles. Whereas these pathways operate under different nutritional conditions, they all employ common machinery with only a few specific factors assisting in the choice of the delivery program and the membrane source for the sequestering vesicle. We found that the YKR020w gene product is essential for Cvt vesicle formation but not for pexophagy or induction of autophagy. Autophagosomes in the ykr020wdelta mutant, however, have a reduced size. We demonstrate that Ykr020 is a subunit of the Vps fifty-three tethering complex, composed of Vps52, Vps53, and Vps54, which is required for retrograde traffic from the early endosome back to the late Golgi, and for this reason we named it Vps51. This complex participates in a fusion event together with Tlg1 and Tlg2, two SNAREs also shown to be necessary for Cvt vesicle assembly. In particular, those factors are essential to correctly target the prApe1-Cvt19-Cvt9 complex to the preautophagosomal structure, the site of Cvt vesicle formation.
自噬、过氧化物酶体自噬和Cvt途径是通过双膜胞质囊泡将水解酶和底物递送至酵母液泡/溶酶体的过程。尽管这些途径在不同的营养条件下发挥作用,但它们都使用共同的机制,只有少数特定因子协助选择递送程序和隔离囊泡的膜来源。我们发现YKR020w基因产物对Cvt囊泡形成至关重要,但对过氧化物酶体自噬或自噬诱导并非必需。然而,ykr020wdelta突变体中的自噬体尺寸减小。我们证明Ykr020是Vps五十三系链复合物的一个亚基,该复合物由Vps52、Vps53和Vps54组成,是从早期内体逆向运输回晚期高尔基体所必需的,因此我们将其命名为Vps51。该复合物与Tlg1和Tlg2一起参与融合事件,这两个SNARE蛋白也被证明是Cvt囊泡组装所必需的。特别是,这些因子对于将prApe1-Cvt19-Cvt9复合物正确靶向自噬前体结构(即Cvt囊泡形成的部位)至关重要。