trans-SNARE complex assembly and yeast vacuole membrane fusion
trans-SNARE complex assembly and yeast vacuole membrane fusion
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DOI：
10.1073/pnas.0702290104
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发表时间：
2007-05-22
影响因子：
11.1
通讯作者：
Wickner, William T.
中科院分区：
文献类型：
--
作者：
Collins, Kevin M.;Wickner, William T.
cis-SNARE 复合物（锚定在一个膜上）被 Sec17p(a-SNAP) 和 Sec18p(NSF) 分解，从而允许未配对的 SNARE 进行反式组装。我们现在报告了酵母液泡对接过程中反式SNARE复合物形成的直接测定。高浓度的 SNARE Vam7p 或 Nyv1p 或添加 HOPS（同型融合和液泡蛋白分选）（一种带有 Secl/Munc18 家族亚基的 Ypt7p (Rab) 效应复合物）可促进 SNARE 复合物的组装和融合。针对 Ypt7p、HOPS 或关键调节脂质的抑制剂可阻止反式 SNARE 复合物组装和随后的融合。引人注目的是，脂质配体 MED（肉豆蔻酰化富含丙氨酸的 C 激酶底物效应结构域）或浓度升高的 Sec17p（可以取代 SNARE 复合物中的 HOPS）允许完全反式 SNARE 配对，但阻止融合。这些发现表明，有效的融合需要反式-SNARE 复合体与 HOPS 等因子以及随后调控的脂质重排相关。
cis-SNARE complexes (anchored in one membrane) are disassembled bySec17p(a-SNAP)and Sec18p(NSF), permitting the unpaired SNAREs to assemble in trans. We now report a direct assay of trans-SNARE complex formation during yeast vacuole docking. SNARE complex assembly and fusion is promoted by high concentrations of the SNARE Vam7p or Nyv1p or by addition of HOPS (homotypic fusion and vacuole protein sorting), a Ypt7p (Rab)effector complex with a Secl/Munc18-family subunit. Inhibitors that target Ypt7p, HOPS, or key regulatory lipids prevent trans-SNARE complex assembly and ensuing fusion. Strikingly, the lipid ligand MED (myristoylated alanine-rich C kinase substrate effector domain) or elevated concentrations of Sec17p, which can displace HOPS from SNARE complexes, permit full trans-SNARE pairing but block fusion. These findings suggest that efficient fusion requires trans-SNARE complex associations with factors such as HOPS and subsequent regulated lipid rearrangements.