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Expression of transfected vimentin genes in differentiating murine erythroleukemia cells reveals divergent cis-acting regulation of avian and mammalian vimentin sequences

机译:转染的波形蛋白基因在分化的鼠红白血病细胞中的表达揭示了禽和哺乳动物波形蛋白序列的顺式作用调控

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摘要

We studied the expression of transfected chicken and hamster vimentin genes in murine erythroleukemia (MEL) cells. MEL cells normally repress the levels of endogenous mouse vimentin mRNA during inducer-mediated differentiation, resulting in a subsequent loss of vimentin filaments. Expression of vimentin in differentiating MEL cells reflects the disappearance of vimentin filaments during mammalian erythropoiesis in vivo. In contrast, chicken erythroid cells express high levels of vimentin mRNA and vimentin filaments during terminal differentiation. We demonstrate here that chicken vimentin mRNA levels increase significantly in differentiating transfected MEL cells, whereas similarly transfected hamster vimentin genes are negatively regulated. In conjunction with in vitro nuclear run-on transcription experiments, these results suggest that the difference in vimentin expression in avian and mammalian erythropoiesis is due to a divergence of cis-linked vimentin sequences that are responsible for transcriptional and posttranscriptional regulation of vimentin gene expression. Transfected chicken vimentin genes produce functional vimentin protein and stable vimentin filaments during MEL cell differentiation, further demonstrating that the accumulation of vimentin filaments is determined by the abundance of newly synthesized vimentin.
机译:我们研究了转染的鸡和仓鼠波形蛋白基因在鼠红细胞白血病(MEL)细胞中的表达。 MEL细胞通常在诱导剂介导的分化过程中抑制内源性小鼠波形蛋白mRNA的水平,导致波形蛋白丝的随后丢失。波形蛋白在分化的MEL细胞中的表达反映了哺乳动物体内红细胞生成过程中波形蛋白丝的消失。相反,鸡红系细胞在终末分化过程中表达高水平的波形蛋白mRNA和波形蛋白细丝。在这里,我们证明了鸡波形蛋白mRNA水平在分化转染的MEL细胞中显着增加,而相似转染的仓鼠波形蛋白基因则受到负调控。结合体外核运行转录实验,这些结果表明,波形蛋白在禽类和哺乳动物红细胞生成中的表达差异是由于顺式连接波形蛋白序列的差异所致,该序列负责波形蛋白基因表达的转录和转录后调控。转染的鸡波形蛋白基因在MEL细胞分化过程中产生功能性波形蛋白蛋白和稳定的波形蛋白细丝,进一步表明波形蛋白细丝的积累取决于新合成的波形蛋白的丰度。

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