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Experimental verification of Myocyte Enchancer Factor 2 (MEF2) and Serum Response Factor (SRF) interaction predicted by computational methods .

机译:通过计算方法预测的肌细胞增强因子2(MEF2)和血清反应因子(SRF)相互作用的实验验证。

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摘要

In this work, a bioinformatic approach for identification of co-occuring transcription factor binding sites is presented. Computational analysis combined with an enhanced method which uses optimized matrix thresholds revealed co-occurrence of binding sites between several transcription factors and Myocyte Enhancer Factor 2 (MEF2). A predicted interaction between MEF2A and Serum Response Factor (SRF) was chosen for further analysis. The muscle creatine kinase (MCK) enhancer contains the Serum Response Element (SRE) and two putative MEF2 binding sites. Co-expression of MEF2A and SRF proteins showed that the MCK enhancer is synergistically activated by this combination of transcription factors. Analysis of the c-jun and c-fos promoters as well as a GAL4- based one hybrid reporter gene assay indicated a requirement of both binding sites for synergistic transcriptional activation. Mutation analysis indicated that "loss of binding" mutations of the MEF2 or SRE potently eradicated the synergistic effect observed on the MCK enhancer. Co-immunoprecipitation experiments revealed a physical interaction between SRF and MEF2 proteins. These results validate the in silico approach of predicting transcription factor interactions and suggest this approach could be a versatile method for prediction of transcription factor interactions.
机译:在这项工作中,提出了一种生物信息学方法,用于鉴定共现的转录因子结合位点。计算分析与使用优化的矩阵阈值的增强方法相结合,揭示了多个转录因子和心肌细胞增强因子2(MEF2)之间结合位点的共存。选择MEF2A和血清反应因子(SRF)之间的预测相互作用进行进一步分析。肌肉肌酸激酶(MCK)增强剂包含血清反应元件(SRE)和两个推定的MEF2结合位点。 MEF2A和SRF蛋白的共表达表明,MCK增强子被转录因子的这种组合协同激活。分析c-jun和c-fos启动子,以及基于GAL4的一种杂交报告基因分析表明,需要两个结合位点才能实现协同转录激活。突变分析表明,MEF2或SRE的“结合缺失”突变有效消除了在MCK增强子上观察到的协同效应。免疫共沉淀实验揭示了SRF和MEF2蛋白之间的物理相互作用。这些结果验证了预测转录因子相互作用的计算机方法,并表明该方法可能是预测转录因子相互作用的通用方法。

著录项

  • 作者

    Pustylnik, Sofia.;

  • 作者单位

    York University (Canada).;

  • 授予单位 York University (Canada).;
  • 学科 Biology Molecular.
  • 学位 M.Sc.
  • 年度 2010
  • 页码 118 p.
  • 总页数 118
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

  • 入库时间 2022-08-17 11:36:51

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