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首页> 外文期刊>Molecular and Cellular Biology >Somatic mRNA Turnover Mutants Implicate Tristetraprolin in the Interleukin-3 mRNA Degradation Pathway
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Somatic mRNA Turnover Mutants Implicate Tristetraprolin in the Interleukin-3 mRNA Degradation Pathway

机译：体细胞mRNA营业额突变体牵涉Tristetraprolin在白介素3 mRNA降解途径中。

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Control of mRNA stability is critical for expression of short-lived transcripts from cytokines and proto-oncogenes. Regulation involves an AU-rich element (ARE) in the 3′ untranslated region (3′UTR) and cognatetrans-acting factors thought to promote either degradation or stabilization of the mRNA. In this study we present a novel approach using somatic cell genetics designed to identify regulators of interleukin-3 (IL-3) mRNA turnover. Mutant cell lines were generated from diploid HT1080 cells transfected with a reporter construct containing green fluorescent protein (GFP) linked to the IL-3 3′UTR. GFP was expressed at low levels due to rapid decay of the mRNA. Following chemical mutagenesis and selection of GFP-overexpressing cells, we could isolate three mutant clones (slowA, slowB, and slowC) with a specific, trans-acting defect in IL-3 mRNA degradation, while the stability of IL-2 and tumor necrosis factor alpha reporter transcripts was not affected. Somatic cell fusion experiments revealed that the mutants are genetically recessive and form two complementation groups. Expression of the tristetraprolin gene in both groups led to reversion of the mutant phenotype, thereby linking this gene to the IL-3 mRNA degradation pathway. The genetic approach described here should allow identification of the defective functions by gene transfer and is also applicable to the study of other mRNA turnover pathways.
机译：mRNA稳定性的控制对于来自细胞因子和原癌基因的短时转录本的表达至关重要。调控涉及3'非翻译区（3'UTR）中的富含AU的元件（ARE）和相关的 trans 作用因子，这些因子被认为可以促进mRNA的降解或稳定。在这项研究中，我们提出了一种使用体细胞遗传学的新颖方法，旨在识别白介素3（IL-3）mRNA转换的调节剂。从二倍体HT1080细胞中产生突变细胞系，该二倍体HT1080细胞已用含有与IL-3 3'UTR连接的绿色荧光蛋白（GFP）的报告基因构建体转染。由于mRNA的快速衰减，GFP以低水平表达。在化学诱变和选择GFP过表达的细胞后，我们可以分离出三个突变克隆（slowA，slowB和slowC），它们在IL-3 mRNA降解中具有特异的 trans 作用缺陷，而稳定性IL-2和肿瘤坏死因子α报告基因的表达不受影响。体细胞融合实验表明，突变体具有遗传隐性，并形成两个互补基团。 Tristetraprolin基因在两组中的表达导致突变表型的逆转，从而使该基因与IL-3 mRNA降解途径相关。此处描述的遗传方法应允许通过基因转移来鉴定功能缺陷，并且还可以用于研究其他mRNA转换途径。

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