Gordon David E, Chia Joanne, Jayawardena Kamburpola, Antrobus Robin, Bard Frederic, Peden Andrew A
University of California San Francisco, Department of Cellular and Molecular Pharmacology, San Francisco, CA, United States of America.
Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore.
PLoS Genet. 2017 Apr 12;13(4):e1006698. doi: 10.1371/journal.pgen.1006698. eCollection 2017 Apr.
The cellular machinery required for the fusion of constitutive secretory vesicles with the plasma membrane in metazoans remains poorly defined. To address this problem we have developed a powerful, quantitative assay for measuring secretion and used it in combination with combinatorial gene depletion studies in Drosophila cells. This has allowed us to identify at least three SNARE complexes mediating Golgi to PM transport (STX1, SNAP24/29 and Syb; STX1, SNAP24/29 and YKT6; STX4, SNAP24 and Syb). RNAi mediated depletion of YKT6 and VAMP3 in mammalian cells also blocks constitutive secretion suggesting that YKT6 has an evolutionarily conserved role in this process. The unexpected role of YKT6 in plasma membrane fusion may in part explain why RNAi and gene disruption studies have failed to produce the expected phenotypes in higher eukaryotes.
后生动物中组成型分泌囊泡与质膜融合所需的细胞机制仍不清楚。为了解决这个问题，我们开发了一种强大的定量测定分泌的方法，并将其与果蝇细胞中的组合基因缺失研究相结合。这使我们能够鉴定出至少三种介导高尔基体到质膜运输的SNARE复合体（STX1、SNAP24/29和Syb；STX1、SNAP24/29和YKT6；STX4、SNAP24和Syb）。RNA干扰介导的哺乳动物细胞中YKT6和VAMP3的缺失也会阻断组成型分泌，这表明YKT6在这一过程中具有进化保守作用。YKT6在质膜融合中的意外作用可能部分解释了为什么RNA干扰和基因破坏研究未能在高等真核生物中产生预期的表型。
