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鸡细胞质β-肌动蛋白在成肌分化过程中的下调不需要基因启动子,但涉及基因的3'端。

The down-regulation of the chicken cytoplasmic beta actin during myogenic differentiation does not require the gene promoter but involves the 3' end of the gene.

作者信息

Lohse P, Arnold H H

机构信息

Department of Toxicology, Medical School, University of Hamburg, FRG.

出版信息

Nucleic Acids Res. 1988 Apr 11;16(7):2787-803. doi: 10.1093/nar/16.7.2787.

Abstract

The chicken cytoplasmic beta actin gene is ubiquitously expressed in all cell types. In terminally differentiated muscle cells, however, the concentration of beta actin specific mRNA is down-regulated to scarcely detectable levels. To test for gene regions which are involved in the muscle specific reduction of beta actin specific mRNA, the isolated complete chicken beta actin gene or chimeric gene constructs containing parts of the gene were stably transfected into the myogenic mouse cell line C2C12 and their transcriptional activity was compared in proliferating myoblasts and postmitotic myotubes. A hybrid construct containing the beta actin promoter fused to the bacterial CAT gene showed high and constitutive expression during myocyte differentiation. In contrast, constructs containing the SV40 early promoter linked to the 3' end of the beta actin gene led to a marked reduction of beta actin transcripts in differentiated C2C12 myotubes. The stability of beta actin mRNA was analyzed in actinomycin D treated cells and found to be virtually unchanged in myotubes as compared to myoblasts. These results suggest that a sequence element located in the 3' end or 3' flanking region of the beta actin gene confers the myotube specific down-regulation that is not primarily due to destabilization of mRNA.

摘要

鸡细胞质β-肌动蛋白基因在所有细胞类型中均普遍表达。然而,在终末分化的肌肉细胞中,β-肌动蛋白特异性mRNA的浓度下调至几乎检测不到的水平。为了检测参与β-肌动蛋白特异性mRNA肌肉特异性减少的基因区域,将分离的完整鸡β-肌动蛋白基因或包含该基因部分的嵌合基因构建体稳定转染到成肌小鼠细胞系C2C12中,并在增殖的成肌细胞和有丝分裂后的肌管中比较它们的转录活性。一个包含与细菌CAT基因融合的β-肌动蛋白启动子的杂交构建体在肌细胞分化过程中表现出高组成型表达。相反,包含与β-肌动蛋白基因3'末端相连的SV40早期启动子的构建体导致分化的C2C12肌管中β-肌动蛋白转录本显著减少。在放线菌素D处理的细胞中分析了β-肌动蛋白mRNA的稳定性,发现与成肌细胞相比,肌管中的稳定性几乎没有变化。这些结果表明,位于β-肌动蛋白基因3'末端或3'侧翼区域的一个序列元件赋予了肌管特异性下调,这主要不是由于mRNA的不稳定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c527/336433/9ccd13c4a548/nar00150-0053-a.jpg

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