Edwards E L, Patrick M H, Ratliff R L, Gray D M
Program in Molecular and Cell Biology, University of Texas, Dallas, Richardson 75083-0688.
Biochemistry. 1990 Jan 23;29(3):828-36. doi: 10.1021/bi00455a033.
Previous experiments have established that in certain synthetic oligomeric DNA sequences, including mixtures of d(AACC)5 with d(CCTT)5, adenine-thymine (A.T) base pairs form to the exclusion of neighboring protonated cytosine-cytosine (C.C+) base pairs [Edwards, E., Ratliff, R., & Gray, D. (1988) Biochemistry 27, 5166-5174]. In the present work, circular dichroism and other measurements were used to study DNA oligomers that represented two additional classes with respect to the formation of A.T and/or C.C+ base pairs. (1) One class included two sets of repeating pentameric DNA sequences, d(CCAAT)3-6 and d(AATCC)4,5. For both of these sets of oligomers, an increase in the magnitude of the long-wavelength positive CD band centered at about 280 nm occurred as the pH was lowered from 7 to 5 at 0.1 and 0.5 M Na+, indicating that C.C+ base pairs formed. Even though it may have been possible for these oligomers to form duplexes with two antiparallel A.T base pairs per pentamer, no A.T base pairing was detected by monitoring the CD changes at 250 nm. Thus, spectral data showed that as few as 40% C.C+ base pairs were stable in two sets of oligomers in which A.T base pairs did not form adjacent to, or in place of, C.C+ base pairs. (2) Another class of oligomer was represented by d(C4A4T4C4), which was studied by CD, HPLC, and centrifugation experiments. We confirmed previous work that this sequence was able to form both types of base pairs as the pH and temperature were lowered [Gray, D., Cui, T., & Ratliff, R. (1984) Nucleic Acids Res. 12, 7565-7580].(ABSTRACT TRUNCATED AT 250 WORDS)
先前的实验已证实,在某些合成寡聚DNA序列中,包括d(AACC)5与d(CCTT)5的混合物,腺嘌呤 - 胸腺嘧啶(A.T)碱基对形成,而相邻的质子化胞嘧啶 - 胞嘧啶(C.C+)碱基对则不形成[爱德华兹,E.,拉特利夫,R.,& 格雷,D.(1988年)《生物化学》27卷,5166 - 5174页]。在本研究中,利用圆二色性及其他测量方法来研究DNA寡聚物,这些寡聚物在A.T和/或C.C+碱基对形成方面代表了另外两类。(1)一类包括两组重复的五聚体DNA序列,d(CCAAT)3 - 6和d(AATCC)4,5。对于这两组寡聚物,当在0.1和0.5 M Na+条件下pH从7降至5时,以约280 nm为中心的长波长正CD带的强度增加,表明形成了C.C+碱基对。尽管这些寡聚物有可能形成每个五聚体有两个反平行A.T碱基对的双链体,但通过监测250 nm处的CD变化未检测到A.T碱基配对。因此,光谱数据表明在两组不形成与C.C+碱基对相邻或替代C.C+碱基对的A.T碱基对的寡聚物中,低至40%的C.C+碱基对是稳定的。(2)另一类寡聚物由d(C4A4T4C4)代表,通过CD、HPLC和离心实验对其进行了研究。我们证实了先前的研究结果,即随着pH和温度降低,该序列能够形成两种类型的碱基对[格雷,D.,崔,T.,& 拉特利夫,R.(1984年)《核酸研究》12卷,7565 - 7580页]。(摘要截于250字)