Apg5p functions in the sequestration step in the cytoplasm-to-vacuole targeting and macroautophagy pathways
Apg5p functions in the sequestration step in the cytoplasm-to-vacuole targeting and macroautophagy pathways
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DOI：
10.1091/mbc.11.3.969
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发表时间：
2000-03-01
影响因子：
3.3
通讯作者：
Klionsky, DJ
中科院分区：
文献类型：
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作者：
George, MD;Baba, M;Klionsky, DJ
细胞质-液泡靶向（Cvt）途径和大自噬是涉及膜重排以在胞质溶胶中形成螯合囊泡，随后将其货物递送到液泡的动态事件。这一过程需要多种蛋白质的协同作用，包括Apg 5 p。最近，它表明，另一种蛋白质所需的，输入氨肽酶I（API）和自噬，Apg 12 p，是共价连接到Apg 5 p通过E1样酶，Apg 7 p的作用。我们已经进行了Apg 5 p功能的分析，以更好地了解这种新的nonubiquitin缀合反应在这些进口途径中的作用。我们已经在Cvt途径中产生了第一个温度敏感突变体，命名为apg 5（ts）。apg 5（ts）菌株中API输入的生化分析证实，Apg 5 p是通过Cvt途径输入API的直接需要。通过分析在apg 5（ts）突变体中阻断的API输入阶段，我们已经确定Apg 5 p参与螯合步骤并且是囊泡形成和/或完成所需的。
The cytoplasm-to-vacuole targeting (Cvt) pathway and macroautophagy are dynamic events involving the rearrangement of membrane to form a sequestering vesicle in the cytosol, which subsequently delivers its cargo to the vacuole. This process requires the concerted action of various proteins, including Apg5p. Recently, it was shown that another protein required for the, import of aminopeptidase I (API) and autophagy, Apg12p, is covalently attached to Apg5p through the action of an E1-like enzyme, Apg7p. We have undertaken an analysis of Apg5p function to gain a better understanding of the role of this novel nonubiquitin conjugation reaction in these import pathways. We have generated the first temperature-sensitive mutant in the Cvt pathway, designated apg5(ts). Biochemical analysis of API import in the apg5(ts) strain confirmed that Apg5p is directly required for the import of API via the Cvt pathway. By analyzing the stage of API import that is blocked in the apg5(ts) mutant, we have determined that Apg5p is involved in the sequestration step and is required for vesicle formation and/or completion.