Hillen W, Wells R D
Nucleic Acids Res. 1980 Nov 25;8(22):5427-44. doi: 10.1093/nar/8.22.5427.
The B goes to A conformational transition caused by high ethanol concentrations was studied for seven DNA restriction fragments with overlapping and known sequences. Since the DNAs are homogeneous and range in GC content from 44-63%, they permit an evaluation of the influence of DNA sequence and base composition on the B goes to A transition. Moreover, their small size (80-301 bp) minimizes precipitation artifacts. The B- form spectra (in low salt) and the transition toward the C- form (in ethanol concentrations below the B goes to A transition) agree with prior measurements on chromosomal DNAs and are similar for all seven DNAs. At higher ethanol concentrations (80%), all fragments undergo a transition to the A- form as judged by the large increase of the positive CD band at 270 nm. Difference spectra among the fragments reveal minor differences between the A- form spectra. The ethanol concentration necessary to cause this transition is 72 +/- 2% for all fragments, thus excluding a preference of the CAP-, E. coli RNA polymerase-, or lac repressor-binding sequences for the A- form. The kinetics of the B goes to A transition in 80% ethanol are biphasic; the initial rapid transition is an intramolecular B goes to A form shift and the slower transition is an aggregation (but not precipitation) of the DNA
对七个具有重叠且已知序列的DNA限制片段,研究了高乙醇浓度引起的B到A构象转变。由于这些DNA是均一的,GC含量在44%-63%之间,它们使得评估DNA序列和碱基组成对B到A转变的影响成为可能。此外,它们的小尺寸(80-301 bp)使沉淀假象最小化。B型光谱(在低盐中)以及向C型的转变(在低于B到A转变的乙醇浓度下)与先前对染色体DNA的测量结果一致,并且对所有七个DNA来说都是相似的。在较高乙醇浓度(80%)下,通过270 nm处正性圆二色带的大幅增加判断,所有片段都发生向A型的转变。片段之间的差异光谱揭示了A型光谱之间的微小差异。导致这种转变所需的乙醇浓度对所有片段来说都是72±2%,因此排除了CAP-、大肠杆菌RNA聚合酶-或乳糖阻遏蛋白结合序列对A型的偏好。在80%乙醇中B到A转变的动力学是双相的;最初的快速转变是分子内B到A形式的转变,较慢的转变是DNA的聚集(但不是沉淀)