Enhancing recombinant protein production in human cell lines with a constitutive transport element and mRNA export proteins
Enhancing recombinant protein production in human cell lines with a constitutive transport element and mRNA export proteins
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利用组成型转运元件和 mRNA 输出蛋白增强人类细胞系中的重组蛋白产量
DOI：
10.1016/j.jbiotec.2011.03.024
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发表时间：
2011
期刊：
影响因子：
4.1
通讯作者：
Masuda S
中科院分区：
文献类型：
--
作者：
Aihara Y;Fujiwara N;Yamazaki T;Kambe T;Nagao M;Hirose Y;Masuda S
最近对细胞核中mRNA成熟过程的研究已经确定了许多参与mRNA转录、加帽、剪接、末端加工和输出的蛋白质。其中，Tap-p15异二聚体作为mRNA输出受体。Tap-p15通过与mRNA上的Aly或SR蛋白缔合，或通过直接与组成型转运元件（CTE）（一种源自D型逆转录病毒的RNA元件）缔合，募集到细胞核中完全加工的mRNA上，其准备输出到细胞质中。含有CTE的mRNA通过与Tap-p15直接缔合而输出到细胞质，即使在mRNA上不存在TAP募集蛋白如Aly或SR蛋白的情况下。在这里，我们表明CTE的使用增强了重组蛋白在人细胞系中的表达。报告蛋白和Tap-p15的共表达也增强了重组蛋白的表达。此外，CTE和Tap-p15的使用协同地进一步增强了重组蛋白的表达。除了Tap-p15，几个Tap-p15募集蛋白，包括Aly和SR蛋白，增强重组蛋白的表达，尽管独立的CTE。将CTE和Tap-p15募集蛋白掺入蛋白表达系统中可用于增加人细胞中的重组蛋白产量。
Recent research into mRNA maturation processes in the nucleus has identified a number of proteins involved in mRNA transcription, capping, splicing, end processing and export. Among them, the Tap-p15 heterodimer acts as an mRNA export receptor. Tap-p15 is recruited onto fully processed mRNA in the nucleus, which is ready for export to the cytoplasm, through associating with Aly or SR proteins on mRNA, or by directly associating with a constitutive transport element (CTE), an RNA element derived from type D retroviruses. mRNA containing a CTE is exported to the cytoplasm by directly associating with Tap-p15, even in the absence of Tap-recruiting proteins such as Aly or SR proteins on the mRNA. Here, we showed that the use of a CTE enhanced the expression of recombinant protein in human cell lines. The co-expression of reporter proteins and Tap-p15 also enhanced recombinant protein expression. Moreover, the use of a CTE and Tap-p15 synergistically further enhanced the recombinant protein expression. In addition to Tap-p15, several Tap-p15-recruiting proteins, including Aly and SR proteins, enhanced recombinant protein expression, albeit independently of the CTE. The incorporation of a CTE and Tap-p15-recruiting proteins into protein expression system is useful to increase recombinant protein yield in human cells.