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首页> 外文期刊>Journal of Molecular Biology >Structural characterization of the 69 nucleotide potato spindle tuber viroid left-terminal domain by NMR and thermodynamic analysis.
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Structural characterization of the 69 nucleotide potato spindle tuber viroid left-terminal domain by NMR and thermodynamic analysis.

机译：69个核苷酸的马铃薯纺锤块茎类病毒左端结构域的结构表征通过NMR和热力学分析。

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摘要

The 69 nucleotide left-terminal domain (T(L)) of the potato spindle tuber RNA viroid (PSTVd) constitutes one of its five structural elements. Due to a twofold complementary sequence repeat, two possible conformations are proposed for the T(L) secondary structure; an elongated-rod and a bifurcated form. In the present study, two T(L) mutants were designed that remove the symmetry of the sequence repeats and ensure that either the bifurcated or the elongated-rod conformation is thermodynamically favored. Imino 1H and 15N resonances were assigned for both mutants and the native T(L) domain based on 1H-1H NOESY and heteronuclear 1H-15N HSQC high-resolution NMR spectra. The NMR secondary structure analysis of all constructs establishes unambiguously the elongated-rod form as the secondary structure of the native T(L) domain. Temperature-gradient gel electrophoresis and UV melting experiments corroborate these results. A combined secondary structure and sequence analysis of T(L) domains of other Pospiviroidae family members indicates that the elongated-rod form is thermodynamically favored for the vast majority of these viroids.
机译：马铃薯纺锤块茎RNA类病毒（PSTVd）的69个核苷酸的左端结构域（T（L））构成其五个结构元件之一。由于双重互补序列重复，对于T（L）二级结构提出了两种可能的构象。细长杆和分叉的形式。在本研究中，设计了两个T（L）突变体，它们消除了重复序列的对称性，并确保热力学上有利于分叉或长杆构型。基于1H-1H NOESY和异核1H-15N HSQC高分辨率NMR光谱，为突变体和天然T（L）域分配了亚氨基1H和15N共振。所有构建体的NMR二级结构分析均明确地确定了细长杆形式为天然T（L）域的二级结构。温度梯度凝胶电泳和紫外熔融实验证实了这些结果。结合的二级结构和其他Pospiviroidae家族成员的T（L）域的序列分析表明，细长杆的形式在热力学上对绝大多数此类类病毒都有利。

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