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Eμ- myc转基因转录增加和mRNA稳定仅使Myc蛋白适度升高。

Increased transcription of the E mu-myc transgene and mRNA stabilisation produce only a modest elevation in Myc protein.

作者信息

Colley S M, Tilbrook P A, Klinken S P

机构信息

Department of Biochemistry, The University of Western Australia, Royal Perth Hospital.

出版信息

Oncogene. 1997 Jun 5;14(22):2735-9. doi: 10.1038/sj.onc.1201113.

Abstract

Mice bearing the E mu-myc transgene, which links the immunoglobulin heavy chain enhancer (E mu) with c-myc, are predisposed to developing B cell lymphomas. Several B lineage cell lines have been isolated from these animals, and some have been converted to macrophages following infection with v-raf. In this study we compared the regulation of myc expression in E mu-myc B lymphoma lines, their macrophage counterparts and other non-myc transformed B cell lines. Nuclear run-on analyses demonstrated that transcription of the transgene was elevated in E mu-myc B cell lines. Moreover, the presence of a 600 bp phiX174 marker in the 3' end of the transgene produced a marked stabilisation of this RNA species. Consequently, steady state myc mRNA levels in the E mu-myc B lymphoma cells were tenfold higher than the macrophage derivatives and non-myc transformed B lineage lines. Despite the considerable difference in myc RNA levels, the E mu-myc B cell lines contained only 30-50% more Myc protein than the other cell lines. This discrepancy between RNA and protein content was not due to increased degradation of the protein as the half life was normal in the transgenic cell lines. These results indicate that both E mu and phiX174 sequences influence transgenic myc expression and that protein levels do not correlate with RNA content in E mu-myc cell lines.

摘要

携带Eμ-myc转基因的小鼠,该转基因将免疫球蛋白重链增强子(Eμ)与c-myc相连,易患B细胞淋巴瘤。已经从这些动物中分离出几种B谱系细胞系,并且其中一些在感染v-raf后已转化为巨噬细胞。在本研究中,我们比较了Eμ-myc B淋巴瘤细胞系、它们的巨噬细胞对应物以及其他非myc转化的B细胞系中myc表达的调控情况。核转录分析表明,转基因在Eμ-myc B细胞系中的转录水平升高。此外,转基因3'端存在600 bp的phiX174标记导致该RNA物种显著稳定。因此,Eμ-myc B淋巴瘤细胞中的稳态myc mRNA水平比巨噬细胞衍生物和非myc转化的B谱系细胞系高十倍。尽管myc RNA水平存在相当大的差异,但Eμ-myc B细胞系中的Myc蛋白仅比其他细胞系多30-50%。RNA和蛋白质含量之间的这种差异并非由于蛋白质降解增加,因为转基因细胞系中的半衰期是正常的。这些结果表明,Eμ和phiX174序列均影响转基因myc的表达,并且在Eμ-myc细胞系中蛋白质水平与RNA含量不相关。

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