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一项基因组筛选将AUT8鉴定为酿酒酵母中自噬所必需的一个新基因。

A genomic screen identifies AUT8 as a novel gene essential for autophagy in the yeast Saccharomyces cerevisiae.

作者信息

Barth H, Thumm M

机构信息

Institut fuer Biochemie, Universitaet Stuttgart, Pfaffenwaldring 55, 70569 Stuttgart, Germany.

出版信息

Gene. 2001 Aug 22;274(1-2):151-6. doi: 10.1016/s0378-1119(01)00614-x.

Abstract

Autophagy is a starvation-induced transport pathway delivering parts of the cytosol into the lysosome (vacuole) for degradation. Autophagy significantly differs from other transport pathways by using double membrane layered transport intermediates. Based on the identification of autophagy genes in Saccharomyces cerevisiae, which served as a pacemaker for higher cells, our mechanistic knowledge of autophagy notably increased over the past few years. We here identify AUT8 as a novel gene essential for autophagy by screening a collection of approximately 5000 yeast deletion strains, each containing a defined deletion in an individual gene. This collection is a result of the world-wide Saccharomyces deletion project and covers the non-essential genes of the whole yeast genome. Homozygous aut8 Delta cells are impaired in maturation of proaminopeptidase I, and they fail to undergo the cell differentiation process of sporulation. The essential function of AUT8 for autophagy is further demonstrated by the lack of accumulation of autophagic vesicles in the vacuoles of aut8 Delta cells starved of nitrogen in the presence of the proteinase B inhibitor phenylmethylsulfonyl fluoride.

摘要

自噬是一种由饥饿诱导的运输途径,它将部分胞质溶胶输送到溶酶体(液泡)中进行降解。自噬通过使用双膜层运输中间体,与其他运输途径显著不同。基于在酿酒酵母中对自噬基因的鉴定,酿酒酵母充当了高等细胞的起搏器,在过去几年里,我们对自噬的机制认识显著增加。我们通过筛选大约5000个酵母缺失菌株的集合,每个菌株在单个基因中含有一个明确的缺失,从而鉴定出AUT8是自噬所必需的一个新基因。这个集合是全球酿酒酵母缺失项目的成果,涵盖了整个酵母基因组的非必需基因。纯合的aut8Δ细胞在氨肽酶I的成熟过程中受损,并且它们无法经历孢子形成的细胞分化过程。在存在蛋白酶B抑制剂苯甲基磺酰氟的情况下,缺乏氮源的aut8Δ细胞的液泡中自噬小泡没有积累,这进一步证明了AUT8对自噬的重要功能。

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