Characterisation of organised smooth endoplasmic reticulum suggests a route towards synthetic compartmentalisation
Characterisation of organised smooth endoplasmic reticulum suggests a route towards synthetic compartmentalisation
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有组织的平滑内质网的表征表明了一条合成区室化的途径
DOI：
10.1101/2022.10.27.514093
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Sandor A
中科院分区：
文献类型：
--
作者：
Sandor A
亚细胞区隔工程是合成生物学的关键挑战之一。在不同的方法中，从新开始建造合成舱是最令人垂涎但也是最困难的选择。重组内质网(ER)，通过引入重组寡聚ER膜驻留蛋白，是建立一个新的隔室的另一个起点。这些蛋白的存在导致内质网的大量扩张和有组织的平滑内质网(OSER)的形成，OSER是一个大的膜性隔室。然而，OSER的特征很差，我们对其对植物潜在生物学的影响的了解有限。在这里，我们描述了一系列OSER间隔区的特征，并展示了诱导多蛋白结构的结构如何影响最终的间隔室形态，面向胞液的反平行寡聚化结构域被证明是触发OSER形成的关键成分。我们表明，虽然OSER保留了与内质网的连接，但对内质网和胞浆都存在扩散障碍。利用高分辨率定量图像分析，我们还表明，这个大隔室的存在并不会扰乱ER网络的其余部分。此外，组成性表达隔室形成多蛋白的转基因拟南芥生长发育正常。这些特性共同表明，OSER可以被开发为一种植物合成生物学工具，用于区隔，结合几种现有策略的好处。只有一种蛋白质结构是诱导其形成所必需的，并且隔室保留了一层分隔膜和对细胞其余部分的扩散屏障。
Engineering of subcellular compartmentalisation is one of synthetic biology’s key challenges. Among different approaches,de novoconstruction of a synthetic compartment is the most coveted but also most difficult option. Restructuring the endoplasmic reticulum (ER), via the introduction of recombinant oligomerising ER-membrane resident proteins, is an alternative starting point for building a new compartment. The presence of such proteins leads to a massive expansion of the ER and the formation of organised smooth endoplasmic reticulum (OSER), a large membranous compartment. However, OSER is poorly characterised and our understanding of its effect on the underlying biology of the plant is limited. Here we characterise a range of OSER compartments and show how the structure of the inducing polyprotein constructs affect the final compartment morphology, with the cytosolic-facing antiparallel oligomerisation domain demonstrated to be an essential component to trigger OSER formation. We show that while OSER retains a connection to the ER, a diffusional barrier exists to both the ER and the cytosol. Using high-resolution quantitative image analysis, we also show that the presence of this large compartment does not disrupt the rest of the ER network. Moreover, transgenicArabidopsisconstitutively expressing the compartment-forming polyproteins grew and developed normally. These properties collectively suggest that OSER could be developed as a plant synthetic biology tool for compartmentalisation, combining the benefits of several existing strategies. Only a single protein construct is necessary to induce its formation, and the compartment retains a delimiting membrane and a diffusional barrier to the rest of the cell.
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DOI：
10.1007/978-1-4939-7389-7
发表时间：
2018
期刊：
The Journal of Physiology
影响因子：
--
作者：
C. Hawes;V. Kriechbaumer
通讯作者：
V. Kriechbaumer
影响因子：
4.5
作者：
Samalova, Marketa;Fricker, Mark;Moore, Ian
通讯作者：
Moore, Ian
影响因子：
19
作者：
Boeynaems S;Alberti S;Fawzi NL;Mittag T;Polymenidou M;Rousseau F;Schymkowitz J;Shorter J;Wolozin B;Van Den Bosch L;Tompa P;Fuxreiter M
通讯作者：
Fuxreiter M
影响因子：
9.5
作者：
I. Sparkes;C. Hawes;L. Frigerio
通讯作者：
I. Sparkes;C. Hawes;L. Frigerio
DOI：
--
发表时间：
2011
期刊：
影响因子：
--
作者：
Murata T;Hasebe M
通讯作者：
Hasebe M