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酵母液泡Qa-SNARE七聚体重复序列上游的一个短区域通过增强SNARE复合体组装促进膜融合。

A short region upstream of the yeast vacuolar Qa-SNARE heptad-repeats promotes membrane fusion through enhanced SNARE complex assembly.

作者信息

Song Hongki, Wickner William

机构信息

Department of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth, Hanover, NH 03755-3844.

Department of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth, Hanover, NH 03755-3844

出版信息

Mol Biol Cell. 2017 Aug 15;28(17):2282-2289. doi: 10.1091/mbc.E17-04-0218. Epub 2017 Jun 21.

DOI:10.1091/mbc.E17-04-0218
PMID:28637767
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5555656/
Abstract

Whereas SNARE (soluble -ethylmaleimide-sensitive factor attachment protein receptor) heptad-repeats are well studied, SNAREs also have upstream N-domains of indeterminate function. The assembly of yeast vacuolar SNAREs into complexes for fusion can be studied in chemically defined reactions. Complementary proteoliposomes bearing a Rab:GTP and either the vacuolar R-SNARE or one of the three integrally anchored Q-SNAREs were incubated with the tethering/SM protein complex HOPS and the two other soluble SNAREs (lacking a transmembrane anchor) or their SNARE heptad-repeat domains. Fusion required a transmembrane-anchored R-SNARE on one membrane and an anchored Q-SNARE on the other. The N-domain of the Qb-SNARE was completely dispensable for fusion. Whereas fusion can be promoted by very high concentrations of the Qa-SNARE heptad-repeat domain alone, at physiological concentrations the Qa-SNARE heptad-repeat domain alone has almost no fusion activity. The 181-198 region of Qa, immediately upstream of the SNARE heptad-repeat domain, is required for normal fusion activity with HOPS. This region is needed for normal SNARE complex assembly.

摘要

虽然可溶性N - 乙基马来酰亚胺敏感因子附着蛋白受体（SNARE）七聚体重复序列已得到充分研究，但SNARE还具有功能不确定的上游N结构域。酵母液泡SNARE组装成用于融合的复合物可以在化学定义的反应中进行研究。将携带Rab:GTP以及液泡R - SNARE或三种整体锚定的Q - SNARE之一的互补蛋白脂质体与拴系/SM蛋白复合物HOPS以及另外两种可溶性SNARE（缺乏跨膜锚定）或其SNARE七聚体重复结构域一起孵育。融合需要一个膜上的跨膜锚定R - SNARE和另一个膜上的锚定Q - SNARE。Qb - SNARE的N结构域对于融合完全是可有可无的。虽然单独的非常高浓度的Qa - SNARE七聚体重复结构域可以促进融合，但在生理浓度下，单独的Qa - SNARE七聚体重复结构域几乎没有融合活性。Qa的181 - 198区域，即紧接在SNARE七聚体重复结构域上游的区域，是与HOPS正常融合活性所必需的。该区域是正常SNARE复合物组装所必需的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/93306ef0ccfb/2282fig8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/9e6f79b2c472/2282fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/88107cafa188/2282fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/1633d4b127eb/2282fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/21948e9aa392/2282fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/729e56d58878/2282fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/353657e20581/2282fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/0ef57a55cff1/2282fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/93306ef0ccfb/2282fig8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/9e6f79b2c472/2282fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/88107cafa188/2282fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/1633d4b127eb/2282fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/21948e9aa392/2282fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/729e56d58878/2282fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/353657e20581/2282fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/0ef57a55cff1/2282fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/088d/5555656/93306ef0ccfb/2282fig8.jpg

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本文引用的文献

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2
Improved reconstitution of yeast vacuole fusion with physiological SNARE concentrations reveals an asymmetric Rab(GTP) requirement.使用生理SNARE浓度改善酵母液泡融合的重组揭示了不对称的Rab（GTP）需求。
Mol Biol Cell. 2016 Aug 15;27(16):2590-7. doi: 10.1091/mbc.E16-04-0230. Epub 2016 Jul 6.
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Chaperoning SNARE assembly and disassembly.
Sci Adv. 2022 Jun 24;8(25):eabo5272. doi: 10.1126/sciadv.abo5272. Epub 2022 Jun 22.
4
Phosphatidylinositol and phosphatidylinositol-3-phosphate activate HOPS to catalyze SNARE assembly, allowing small headgroup lipids to support the terminal steps of membrane fusion.磷脂酰肌醇和磷脂酰肌醇-3-磷酸激活 HOPS 以催化 SNARE 组装，使小头部脂质能够支持膜融合的最后步骤。
Mol Biol Cell. 2021 Nov 1;32(21):ar19. doi: 10.1091/mbc.E21-07-0373. Epub 2021 Sep 8.
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Sec17/Sec18 can support membrane fusion without help from completion of SNARE zippering.Sec17/Sec18 可以在没有 SNARE 拉链完成帮助的情况下支持膜融合。
Elife. 2021 May 4;10:e67578. doi: 10.7554/eLife.67578.
6
Asymmetric Rab activation of vacuolar HOPS to catalyze SNARE complex assembly.液泡 HOPS 通过非对称 Rab 激活来催化 SNARE 复合物组装。
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7
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J Biol Chem. 2013 Oct 4;288(40):28557-66. doi: 10.1074/jbc.M113.484410. Epub 2013 Aug 16.