Gottesfeld J M, Blanco J, Tennant L L
Department of Molecular Biology, Research Institute of Scripps, La Jolla, California 92037.
Nature. 1987;329(6138):460-2. doi: 10.1038/329460a0.
Rhodes and Klug have recently proposed that the internal control region of the Xenopus 5S RNA gene adopts an A-type DNA structure in solution. This suggestion was based on a Fourier analysis of both the spacing of DNase I cutting sites and on the distribution of G residues in the DNA sequence. Both revealed a approximately 5.6-5.7-base periodicity which the authors interpreted as a structural repeat every half helical turn of A-type DNA. This contention was strengthened by the finding that a 9-base-pair (bp) double-stranded deoxyoligonucleotide corresponding to residues +81 to +89 of the 5S gene exhibits an A' RNA-like crystal structure. This region of DNA is of special interest as it forms the binding site for the 5S gene-specific transcription factor IIIA (TFIIIA). TFIIIA is a Zn2+-binding protein which interacts with both the internal control region of the gene and the 5S transcript. As base-paired regions of RNA are of the A type, it was reasonable to postulate that 5S DNA might also adopt this conformation. We report here that the circular dichroism (CD) spectrum of a synthetic 54-bp deoxyoligonucleotide corresponding to the TFIIIA binding site is similar to the CD spectrum of B-form DNA in solution. Further, DNA-TFIIIA complexes show an unaltered DNA CD component indicating no gross alteration in DNA structure on protein binding.
罗兹和克卢格最近提出,非洲爪蟾5S RNA基因的内部控制区在溶液中采用A型DNA结构。这一观点基于对DNA酶I切割位点间距以及DNA序列中G残基分布的傅里叶分析。两者都揭示了大约5.6 - 5.7个碱基的周期性,作者将其解释为A型DNA每半个螺旋圈的结构重复。对应于5S基因+81至+89位残基的一个9碱基对(bp)的双链脱氧寡核苷酸呈现出类似A' RNA的晶体结构,这一发现进一步支持了上述观点。DNA的这一区域特别引人关注,因为它形成了5S基因特异性转录因子IIIA(TFIIIA)的结合位点。TFIIIA是一种锌结合蛋白,它与基因的内部控制区以及5S转录本相互作用。由于RNA的碱基配对区域是A型的,因此推测5S DNA也可能采用这种构象是合理的。我们在此报告,对应于TFIIIA结合位点的一个合成54 bp脱氧寡核苷酸的圆二色性(CD)光谱与溶液中B型DNA的CD光谱相似。此外,DNA - TFIIIA复合物显示出未改变的DNA CD成分,表明蛋白质结合后DNA结构没有发生总体改变。