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d(GT)n.d(GA)n序列的结构:平行链双链DNA的形成

Structure of d(GT)n.d(GA)n sequences: formation of parallel stranded duplex DNA.

作者信息

Germann M W, Kalisch B W, van de Sande J H

机构信息

Kimmel Cancer Institute, Department of Microbiology and Immunology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.

出版信息

Biochemistry. 1998 Sep 15;37(37):12962-70. doi: 10.1021/bi9726842.

Abstract

Alternating polypurine sequences exhibit remarkable polymorphism. In this study, we report that dGA.dGT sequences form parallel stranded duplex DNA at neutral pH. Using two model hairpins, 3'-d(GT)3-5'5'-T4(AG)3-3' (I) and 3'-d(GT)4-5'5'-T4(AG)4-3' (II), containing 5'5' linkages which direct parallel strand formation, we systematically explored the spectroscopic and thermodynamic properties of parallel stranded d(GA)n.d(GT)n. The parallel stranded hairpins are remarkably stable structures with TM's of 41.5 and 47.5 degreesC (in 0.4 M NaCl) for the shorter and longer hairpins, respectively. The van't Hoff enthalpies of 80.7 and 114 kJ mol-1 are relatively low but are comparable to a parallel stranded d(GA)n duplex. On the basis of the spectroscopic and electrophoretic characteristics, we conclude that parallel strand formation is not restricted to hairpin systems, but also readily occurs in unconstrained dimeric duplexes with the appropriate sequence homologies. Both melting curves and electrophoretic analyses of parallel stranded heteroduplexes in which the sequence enforces specific base pairing demonstrate that G-G and A-T base pairs are formed in d(GA)n.d(GT)n segments.

摘要

交替的多聚嘌呤序列表现出显著的多态性。在本研究中,我们报道了dGA.dGT序列在中性pH条件下形成平行链双链DNA。使用两个模型发夹结构,即3'-d(GT)3-5'5'-T4(AG)3-3'(I)和3'-d(GT)4-5'5'-T4(AG)4-3'(II),它们含有5'5'连接以指导平行链的形成,我们系统地研究了平行链d(GA)n.d(GT)n的光谱和热力学性质。对于较短和较长的发夹结构,平行链发夹分别是非常稳定的结构,在0.4 M NaCl中,其解链温度分别为41.5和47.5摄氏度。80.7和114 kJ mol-1的范特霍夫焓相对较低,但与平行链d(GA)n双链相当。基于光谱和电泳特征,我们得出结论,平行链的形成不限于发夹系统,在具有适当序列同源性的无约束二聚体双链中也很容易发生。对平行链异源双链的熔解曲线和电泳分析表明,在d(GA)n.d(GT)n片段中形成了G-G和A-T碱基对,其中序列强制形成特定的碱基配对。

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