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首页> 外文期刊>The FEBS journal >C -> G base mutations in the CArG box of c-fos serum response element alter its bending flexibility
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C -> G base mutations in the CArG box of c-fos serum response element alter its bending flexibility

机译：c-fos血清反应元件CArG盒中的C-> G碱基突变改变了其弯曲弹性

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摘要

By binding to the CArG box sequence, the serum response factor (SRF) activates several muscle-specific genes, as well as genes that respond to mitogens. The core domain of the SRF (core-SRF) binds as a dimer to the CArG box C(-5)C(-4)A(-3)T(-2)A(-1)T(+1)T(+2)A(+3)G(+4)G(+5) of the c-fos serum response element (SREfos). However, previous studies using 20-mer DNAs have shown that the binding stoichiometry of core-SRF is significantly altered by mutations C-5 -> G (SREGfos) and C-5C-4 -> GG (SREGGfos) of the CArG box [A Huet, A Parlakian, M-C Arnaud, J-M Glandieres, P Valat, S Fermandjian, D Paulin, B Alpert & C Zentz (2005) FEBS J272, 3105-3119]. To understand these effects, we carried out a comparative analysis of the three 20-mer DNAs SREfos, SREGfos and SREGGfos in aqueous solution. Their CD spectra were of the B-DNA type with small differences generated by variations in the mutual arrangement of the base pairs. Analysis by singular value decomposition of a set of Raman spectra recorded as a function of temperature, revealed a premelting transition associated with a conformational shift in the DNA double helices from a bent to a linear form. Time-resolved fluorescence anisotropy shows that the fluorescein reporter linked to the oligonucleotide 5'-ends experiences twisting motions of the double helices related to the interconversion between bent and linear conformers. The three SREs present various bent populations submitted, however, to particular internal dynamics, decisive for the mutual adjustment of binding partners and therefore specific complex formation.
机译：通过与CArG盒序列结合，血清反应因子（SRF）激活了几个肌肉特异性基因，以及对有丝分裂原有反应的基因。 SRF的核心域（core-SRF）作为二聚体与CArG框C（-5）C（-4）A（-3）T（-2）A（-1）T（+1）T结合c-fos血清反应元件（SREfos）的（+2）A（+3）G（+4）G（+5）。但是，先前使用20-mer DNA的研究表明，CArG框的C-5-> G（SREGfos）和C-5C-4-> GG（SREGGfos）突变显着改变了核心SRF的结合化学计量[ Huet，Parlakian，MC Arnaud，JM Glandieres，P Valat，S Fermandjian，D Paulin，B Alpert和C Zentz（2005）FEBS J272，3105-3119]。为了了解这些影响，我们对水溶液中的三个20-mer DNA SREfos，SREGfos和SREGGfos进行了比较分析。它们的CD光谱是B-DNA类型的，由碱基对相互排列的变化产生的微小差异。通过对一组记录为温度的拉曼光谱进行奇异值分解分析，发现与DNA双螺旋结构从弯曲到线性的构象位移有关的预熔转变。时间分辨的荧光各向异性表明，连接至寡核苷酸5'-末端的荧光素报告基因经历了双螺旋的扭曲运动，这与弯曲和线性构象异构体之间的相互转化有关。这三个SREs呈现出不同的倾向群体，但是，它们受到特定的内部动力的影响，对于相互调节结合配偶体并因此形成特定的复合物起决定性作用。

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