Biology and Biological Engineering,Chalmers University of Technology,Gothenburg,Sweden.
Department of Chemistry and Molecular Biology,Gothenburg University,Gothenburg,Sweden.
Q Rev Biophys. 2017 Jan;50:e11. doi: 10.1017/S0033583517000099.
We have discovered a well-defined extended conformation of double-stranded DNA, which we call Σ-DNA, using laser-tweezers force-spectroscopy experiments. At a transition force corresponding to free energy change ΔG = 1·57 ± 0·12 kcal (mol base pair)-1 60 or 122 base-pair long synthetic GC-rich sequences, when pulled by the 3'-3' strands, undergo a sharp transition to the 1·52 ± 0·04 times longer Σ-DNA. Intriguingly, the same degree of extension is also found in DNA complexes with recombinase proteins, such as bacterial RecA and eukaryotic Rad51. Despite vital importance to all biological organisms for survival, genome maintenance and evolution, the recombination reaction is not yet understood at atomic level. We here propose that the structural distortion represented by Σ-DNA, which is thus physically inherent to the nucleic acid, is related to how recombination proteins mediate recognition of sequence homology and execute strand exchange. Our hypothesis is that a homogeneously stretched DNA undergoes a 'disproportionation' into an inhomogeneous Σ-form consisting of triplets of locally B-like perpendicularly stacked bases. This structure may ensure improved fidelity of base-pair recognition and promote rejection in case of mismatch during homologous recombination reaction. Because a triplet is the length of a gene codon, we speculate that the structural physics of nucleic acids may have biased the evolution of recombinase proteins to exploit triplet base stacks and also the genetic code.
我们使用激光镊子力谱实验发现了一种定义明确的双链 DNA 扩展构象,我们称之为 Σ-DNA。在对应于自由能变化 ΔG = 1·57 ± 0·12 kcal(摩尔碱基对)-1 的过渡力下,当用 3'-3'链拉动时,长 60 或 122 个碱基对的富含 GC 的合成序列会急剧转变为 1·52 ± 0·04 倍长的 Σ-DNA。有趣的是,在与重组酶蛋白(如细菌 RecA 和真核 Rad51)形成的 DNA 复合物中也发现了相同程度的延伸。尽管重组反应对于所有生物的生存、基因组维护和进化都至关重要,但它在原子水平上仍未被理解。我们在此提出,Σ-DNA 所代表的结构扭曲是核酸固有的,与重组蛋白如何介导序列同源性的识别和执行链交换有关。我们的假设是,均匀拉伸的 DNA 会发生“歧化”,形成由局部 B 型垂直堆叠碱基组成的非均匀 Σ 型。这种结构可能确保了碱基对识别的更高保真度,并在同源重组反应中出现错配时促进其排斥。由于三联体是基因密码子的长度,我们推测核酸的结构物理学可能影响了重组酶蛋白利用三联体碱基堆叠和遗传密码的进化。