McLean M J, Blaho J A, Kilpatrick M W, Wells R D
Proc Natl Acad Sci U S A. 1986 Aug;83(16):5884-8. doi: 10.1073/pnas.83.16.5884.
The capacity of six sequences with different numbers and orientations of A.T pairs flanked by alternating C.G pairs to adopt left-handed structures was evaluated in recombinant plasmids. A series of synthetic oligodeoxynucleotides were cloned into the BamHI site of pRW790, a small plasmid (approximately 2 kilobases) prepared especially for conformational studies of this type. Supercoil relaxation studies by two-dimensional gel electrophoresis on topoisomers of each plasmid revealed the energetics and structures of the left-handed helices. Also, the presence of supercoil-induced altered DNA conformations within the inserts of topoisomer populations of the plasmids was detected by reaction with S1 nuclease followed by restriction mapping of the cleavage sites. We conclude that consecutive T.A base pairs, whether alternating (TATA) or contiguous (TTTT), can adopt a left-handed conformation (presumably Z) when flanked by reasonably short runs of alternating (C-G)n (n = 3-5). Thus, these results substantially broaden the range of DNA sequences that can adopt left-handed Z conformations.
在重组质粒中评估了六种两侧由交替的C.G对侧翼的具有不同数量和方向的A.T对序列形成左手结构的能力。一系列合成寡脱氧核苷酸被克隆到pRW790的BamHI位点,pRW790是一种特别为这类构象研究制备的小质粒(约2千碱基)。通过二维凝胶电泳对每个质粒的拓扑异构体进行超螺旋松弛研究,揭示了左手螺旋的能量学和结构。此外,通过与S1核酸酶反应,随后对切割位点进行限制性图谱分析,检测了质粒拓扑异构体群体插入片段内超螺旋诱导的DNA构象变化。我们得出结论,当两侧有相当短的交替(C-G)n(n = 3-5)片段时,连续的T.A碱基对,无论是交替的(TATA)还是相邻的(TTTT),都可以形成左手构象(推测为Z型)。因此,这些结果大大拓宽了能够形成左手Z构象的DNA序列范围。