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Transcriptional and posttranscriptional regulation of transcription factor expression in Arabidopsis roots

机译:拟南芥根中转录因子表达的转录和转录后调控

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Understanding how the expression of transcription factor (TF) genes is modulated is essential for reconstructing gene regulatory networks. There is increasing evidence that sequences other than upstream noncoding can contribute to modulating gene expression, but how frequently they do so remains unclear. Here, we investigated the regulation of TFs expressed in a tissue-enriched manner in Arabidopsis roots. For 61 TFs, we created GFP reporter constructs driven by each TF's upstream noncoding sequence (including the 5'UTR) fused to the GFP reporter gene alone or together with the TF's coding sequence. We compared the visually detectable GFP patterns with endogenous mRNA expression patterns, as defined by a genome-wide microarray root expression map. An automated image analysis method for quantifying GFP signals in different tissues was developed and used to validate our visual comparison method. From these combined analyses, we found that (i) the upstream noncoding sequence was sufficient to recapitulate the mRNA expression pattern for 80% (35/44) of the TFs, and (ii) 25% of the TFs undergo posttranscriptional regulation via microRNA-mediated mRNA degradation (2/24) or via intercellular protein movement (6/24). The results suggest that, for Arabidopsis TFs, upstream noncoding sequences are major contributors to mRNA expression pattern establishment, but modulation of transcription factor protein expression pattern after transcription is relatively frequent. This study provides a systematic overview of regulation of TF expression at a cellular level.
机译:了解转录因子(TF)基因的表达是如何被调节的对于重建基因调控网络至关重要。越来越多的证据表明,除了上游非编码序列以外的其他序列也可以参与调节基因表达,但目前尚不清楚它们的执行频率。在这里,我们调查了拟南芥根中以组织富集方式表达的TF的调控。对于61个TF,我们创建了由每个TF上游非编码序列(包括5'UTR)驱动的GFP报告基因构建体,该非编码序列单独或与TF的编码序列融合到GFP报告基因上。我们将视觉上可检测到的GFP模式与内源性mRNA表达模式进行了比较,后者由全基因组微阵列根表达图定义。开发了一种用于量化不同组织中GFP信号的自动图像分析方法,并用于验证我们的视觉比较方法。从这些组合分析中,我们发现(i)上游非编码序列足以概括80%(35/44)TF的mRNA表达模式,并且(ii)25%TF通过microRNA-进行转录后调控介导的mRNA降解(2/24)或通过细胞间蛋白移动(6/24)。结果表明,对于拟南芥TF,上游非编码序列是mRNA表达模式建立的主要贡献者,但是转录后转录因子蛋白质表达模式的调节相对频繁。这项研究提供了在细胞水平上调节TF表达的系统概述。

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