Miller P S, Bhan P, Cushman C D, Trapane T L
Department of Biochemistry, School of Hygiene and Public Health, Johns Hopkins University, Baltimore, Maryland 21205.
Biochemistry. 1992 Jul 28;31(29):6788-93. doi: 10.1021/bi00144a020.
Two oligodeoxyribonucleotides, d-CTTCTTTTTTATTTT, I(A), and d-ATTATTTTTTATTTT, II(A), where C is 5-methylcytosine and A is 8-oxoadenine, were prepared and their interactions with the duplex d-GAAGAAAAAAYAAAA/d-TTTTZTTTTTTCTTC, III.IV(Y.Z), were studied. Oligomers I(A) and II(A) each form triplexes with III.IV(G.C) at temperatures below 20 degrees C as shown by continuous variation experiments, melting experiments, and circular dichroism (CD) spectroscopy. The CD spectra of these triplexes are almost identical to those formed by I(C) and II(C), oligomers which contain cytosine in place of 8-oxoadenine. This suggests that the 8-oxoadenine-containing triplexes have conformations which are very similar to those of the cytosine-containing triplexes. The melting temperature (Tm) for dissociation of the third strand of triplex II.III.IV(A.G.C) is 22 degrees C at pH 7.0 and 8.0, whereas the Tm of the corresponding transition in triplex II.III.IV(C.G.C) decreases from 28 degrees C at pH 7.0 to 17 degrees C at pH 8.0. The pH dependence of the Tm in the latter triplex reflects the necessity of protonating the N-3 of cytosine in order for it to form two hydrogen bonds with G of the G.C base pair. It appears that the keto form of 8-oxoadenine can potentially form two hydrogen bonds with the N-7 and O-6 atoms of G of the G.C base pair, when the 8-oxoadenine is in the syn conformation and in contrast to cytosine does not require protonation of the base. Oligomer I(A) does not form triplexes with III.IV(Y.Z) when Y.Z is A.T or T.A.(ABSTRACT TRUNCATED AT 250 WORDS)
制备了两条寡脱氧核糖核苷酸,即d-CTTCTTTTTTATTTT,I(A),和d-ATTATTTTTTATTTT,II(A),其中C为5-甲基胞嘧啶,A为8-氧代腺嘌呤,并研究了它们与双链体d-GAAGAAAAAAYAAAA/d-TTTTZTTTTTTCTTC,III.IV(Y.Z)的相互作用。如连续变化实验、熔解实验和圆二色性(CD)光谱所示,寡聚物I(A)和II(A)在温度低于20摄氏度时均与III.IV(G.C)形成三链体。这些三链体的CD光谱与由I(C)和II(C)形成的光谱几乎相同,I(C)和II(C)是用胞嘧啶取代8-氧代腺嘌呤的寡聚物。这表明含8-氧代腺嘌呤的三链体具有与含胞嘧啶的三链体非常相似的构象。三链体II.III.IV(A.G.C)第三条链解离的熔解温度(Tm)在pH 7.0和8.0时为22摄氏度,而三链体II.III.IV(C.G.C)相应转变的Tm从pH 7.0时的28摄氏度降至pH 8.0时的17摄氏度。后一种三链体中Tm的pH依赖性反映了胞嘧啶的N-3质子化以使其与G.C碱基对的G形成两个氢键的必要性。当8-氧代腺嘌呤处于顺式构象时,其酮式可能与G.C碱基对的G的N-7和O-6原子形成两个氢键,与胞嘧啶不同,它不需要碱基质子化。当Y.Z为A.T或T.A.时,寡聚物I(A)不与III.IV(Y.Z)形成三链体。(摘要截于250字)