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鉴定参与转录/翻译的蛋白质(eEF1A1)作为 Bax 诱导的细胞凋亡抑制剂。

Identification of proteins involved in transcription/translation (eEF 1A1) as an inhibitor of Bax induced apoptosis.

机构信息

School of Life Sciences, Medical School, University of Nottingham, Nottingham, NG7 2UH, UK.

Leicester School of Pharmacy, De Montfort University, Leicester, LE1 9BH, UK.

出版信息

Mol Biol Rep. 2020 Sep;47(9):6785-6792. doi: 10.1007/s11033-020-05736-5. Epub 2020 Sep 1.

Abstract

Eukaryotic elongation factor 1A1 (eEF1A1) is central to translational activity. It is involved in complexes that form signal transduction with protein kinase C, as well as being a signal transducer and activator of transcription 3. eEF1A1 and eEF1A2 are isoforms of the alpha subunit of elongating factor 1 complex. It has been reported that eEF1A1 is expressed in most human tissues but the brain, skeletal muscle and heart. eEF1A1 has been linked to both apoptosis and anti-apoptotic activities. In this study, eEF1A1 was co-expressed with Bax, a proapoptotic protein via heterologous expression of recombinant DNA in yeast cells. Assays were carried out to monitor the fate and state of yeast cells when eEF1A1 was co-expressed with Bax. The yeast strain (bearing an integrated copy of the Bax gene) was transformed with an episomal 2-micron plasmid that encodes HA-tagged eEF1A1 gene. The resultant strain would allow co-expression of Bax and eEF1A1 in yeast cells, Bax being under the control of the GAL1 promoter, while the PGK1 promoter drives eEF1A1 expression. Bcl 2A1, a known anti-apoptotic protein, was also co-expressed with Bax in yeast cells as a positive control, to study the anti-apoptotic characteristic of eEF-1A1. The part eEF1A1 plays in apoptosis has been contentious, amidst the pro and anti-apoptotic properties of eEF1A1, it was shown clearly, in this study that eEF1A1 portrays only anti-apoptotic property in the presence of pro-apoptotic protein, Bax.

摘要

真核延伸因子 1A1(eEF1A1)是翻译活性的核心。它参与与蛋白激酶 C 形成信号转导的复合物,并且是信号转导子和转录激活子 3 的信号转导子和转录激活子。eEF1A1 和 eEF1A2 是延伸因子 1 复合物的α亚基同工型。据报道,eEF1A1 在大多数人体组织中表达,但在脑、骨骼肌和心脏中不表达。eEF1A1 与细胞凋亡和抗细胞凋亡活性有关。在这项研究中,eEF1A1 通过在酵母细胞中异源表达重组 DNA 与促凋亡蛋白 Bax 共表达。进行了测定以监测当 eEF1A1 与 Bax 共表达时酵母细胞的命运和状态。该酵母株(携带 Bax 基因的整合拷贝)被转化为携带编码 HA 标记的 eEF1A1 基因的附加体 2 微米质粒。该菌株将允许 Bax 和 eEF1A1 在酵母细胞中共表达,Bax 受 GAL1 启动子的控制,而 PGK1 启动子驱动 eEF1A1 表达。Bcl 2A1,一种已知的抗凋亡蛋白,也与 Bax 在酵母细胞中共表达作为阳性对照,以研究 eEF-1A1 的抗凋亡特性。eEF1A1 在细胞凋亡中的作用一直存在争议,在 eEF1A1 的促凋亡和抗凋亡特性中,本研究清楚地表明,在存在促凋亡蛋白 Bax 的情况下,eEF1A1 仅表现出抗凋亡特性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7bb7/7561549/7a2a65bc1c21/11033_2020_5736_Fig1_HTML.jpg

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