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首页> 外文期刊>European Journal of Cell Biology: Journal of Deutsche Gesellschaft fur Elektronenmikroskopie: Journal of Deutsche Gesellschaft fur Zellbiologie >Members of a mammalian SNARE complex interact in the endoplasmic reticulum in vivo and are found in COPI vesicles.
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Members of a mammalian SNARE complex interact in the endoplasmic reticulum in vivo and are found in COPI vesicles.

机译：哺乳动物SNARE复合体的成员在体内内质网中相互作用，并在COPI囊泡中发现。

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摘要

Retrograde traffic between the Golgi apparatus and the endoplasmic reticulum (ER) is largely mediated by COPI-coated transport vesicles. In mammalian cells, retrograde traffic can pass through an intermediate compartment. Here, we report that the mammalian soluble N-ethylmaleimide-sensitive factor (NSF) attachment receptor (SNARE) proteins mSec22b, mUse1/D12, mSec20/BNIP1, and syntaxin 18 form a quaternary SNARE complex. Fluorescence resonance energy transfer (FRET) experiments prove that these interactions occur in the ER of living cells. In addition, mUse1/D12 and mSec20/BNIP1 form homo-oligomers in vivo. Furthermore, we show that mSec22b, mUse1/D12, mSec20/BNIP1, and syntaxin 18 are recruited into COPI-coated vesicles formed in vitro. Immunogold electron microscopy confirmed that these SNARE proteins colocalize with the KDEL receptor ERD2 in COPI-coated vesicles. Moreover, both FRET and immunoprecipitation experiments reveal interactions of these SNAREs with both ERD2 and COPI subunits. We conclude that the SNAREs described here are sorted via interaction with components of the COPI-dependent budding complex into Golgi-to-ER retrograde COPI vesicles and function in retrograde transport from the Golgi to the ER Golgi intermediate compartment (ERGIC) or the ER.
机译：高尔基体和内质网（ER）之间的逆行交通很大程度上是由COPI涂层的运输小泡介导的。在哺乳动物细胞中，逆行交通可以通过中间隔室。在这里，我们报告哺乳动物可溶性N-乙基马来酰亚胺敏感因子（NSF）附着受体（SNARE）蛋白mSec22b，mUse1 / D12，mSec20 / BNIP1和syntaxin 18形成了四级SNARE复合物。荧光共振能量转移（FRET）实验证明，这些相互作用发生在活细胞的ER中。此外，mUse1 / D12和mSec20 / BNIP1在体内形成同源寡聚体。此外，我们表明mSec22b，mUse1 / D12，mSec20 / BNIP1和syntaxin 18被招募到体外形成的COPI涂层囊泡中。免疫金电子显微镜检查证实，这些SNARE蛋白与CODEL包被的囊泡中的KDEL受体ERD2共定位。此外，FRET和免疫沉淀实验均揭示了这些SNARE与ERD2和COPI亚基的相互作用。我们得出的结论是，此处描述的SNARE是通过与COPI依赖的芽接复合物的成分相互作用而分类成高尔基体到ER的逆行COPI囊泡，并在从高尔基体到ER高尔基体中隔室（ERGIC）或ER的逆行转运中起作用。

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