Yeast transformation efficiency is enhanced by TORC1- and eisosome-dependent signaling.

Yeast transformation efficiency is enhanced by TORC1- and eisosome-dependent signaling.
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DOI:
10.1002/mbo3.730
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发表时间:
2019-05
期刊:
影响因子:
3.4
通讯作者:
Spanu PD
Spanu PD
中科院分区:
生物学3区
文献类型:
--
作者:
Yu SC;Kuemmel F;Skoufou-Papoutsaki MN;Spanu PD

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面包酵母(Saccharmyces Cerevisiae)的转化在几种实验技术中起着关键作用,但转化的分子机制尚不清楚。在生长和转化培养基中添加氨基酸可以提高转化效率。在这里,我们表明,雷帕霉素复合体1(TORC1)的靶标被氨基酸激活,通过泛素介导的内吞作用促进转化。我们创造了TORC1途径、α-阻滞素和Eisosome相关基因的突变。我们的结果表明,TORC1-Npr1-Art1/Rsp5途径调控酵母转化。根据我们之前的研究,激活这一途径可以使转化效率提高200倍或更多。此外,我们认为DNA可能被胞膜中CAN1(MCC)的结构域所占据。酵母转化的研究可以作为一个平台来了解哺乳动物系统中DNA摄取的机制,这对于优化基因治疗具有临床意义。
Transformation of baker's yeast (Saccharomyces cerevisiae) plays a key role in several experimental techniques, yet the molecular mechanisms underpinning transformation are still unclear. The addition of amino acids to the growth and transformation medium increases transformation efficiency. Here, we show that target of rapamycin complex 1 (TORC1) activated by amino acids enhances transformation via ubiquitin‐mediated endocytosis. We created mutants of the TORC1 pathway, alpha‐arrestins, and eisosome‐related genes. Our results demonstrate that the TORC1‐Npr1‐Art1/Rsp5 pathway regulates yeast transformation. Based on our previous study, activation of this pathway results in up to a 200‐fold increase in transformation efficiency, or greater. Additionally, we suggest DNA may be taken up by domains at the membrane compartment of Can1 (MCC) in the plasma membrane formed by eisosomes. Yeast studies on transformation could be used as a platform to understand the mechanism of DNA uptake in mammalian systems, which is clinically relevant to optimize gene therapy.
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