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首页> 外文期刊>Nucleic acids research >Myosin light chain 1 atrial isoform (MLC1A) is expressed in pre‐B cells under control of the BOB.1/OBF.1 coactivator
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Myosin light chain 1 atrial isoform (MLC1A) is expressed in pre‐B cells under control of the BOB.1/OBF.1 coactivator

机译:肌球蛋白轻链1心房同工型(MLC1A)在BOB.1 / OBF.1共激活因子的控制下在pre-B细胞中表达

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摘要

The BOB.1/OBF.1 protein is a B‐cell‐specific coactivator of the Oct1 and Oct2 transcription factors. It is involved in mediating the transcriptional activity of the Oct proteins. However, animals deficient for BOB.1/OBF.1 showed virtually normal expression of genes that contain octamer motifs in their regulatory regions. To identify new genes that are regulated by BOB.1/OBF.1, we took advantage of a previously described cell system. RNAs differentially expressed in a BOB.1/OBF.1‐deficient pre‐B cell line and a derivative of this cell line expressing a hormone dependent BOB.1/OBF.1‐estrogene receptor (BobER) fusion protein were isolated. Using the cDNA representational difference analysis method we could identify myosin light chain 1 atrial (MLC1A) isoform as a gene regulated by BOB.1/OBF.1. MLC1A was so far unknown to be expressed in tissues other than muscle. Here we demonstrate that MLC1A is indeed expressed in mouse pre‐B cells. Analysis of the expressed mRNA revealed an alternative 5′ promoter element and an alternative splice product, which had not yet been described for the murine gene. Cotransfection experiments with reporter constructs driven by the MLC1A promoter suggest that the regulation by BOB.1/OBF.1 is indirect. Consistent with this conclusion is the observation that transcriptional induction of the endogenous MLC1A gene by BOB.1/OBF.1 requires de novo protein synthesis.
机译:BOB.1 / OBF.1蛋白是Oct1和Oct2转录因子的B细胞特异性共激活因子。它参与介导Oct蛋白的转录活性。但是,缺乏BOB.1 / OBF.1的动物实际上在其调控区中含有八聚体基序的基因正常表达。为了鉴定受BOB.1 / OBF.1调控的新基因,我们利用了先前描述的细胞系统。分离出在BOB.1 / OBF.1缺陷的pre-B细胞系中差异表达的RNA及其表达激素依赖性BOB.1 / OBF.1雌激素受体(BobER)融合蛋白的该细胞系的衍生物。使用cDNA代表性差异分析方法,我们可以鉴定肌球蛋白轻链1心房(MLC1A)同工型为BOB.1 / OBF.1调控的基因。迄今为止,MLC1A尚未在肌肉以外的组织中表达。在这里,我们证明了MLC1A确实在小鼠pre-B细胞中表达。对表达的mRNA的分析显示了替代5'启动子元件和替代剪接产物,尚未针对鼠基因进行描述。与MLC1A启动子驱动的报告基因构建体的共转染实验表明,BOB.1 / OBF.1的调控是间接的。与此结论相一致的是,观察到BOB.1 / OBF.1对内源性MLC1A基因的转录诱导需要从头合成蛋白质。

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