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首页> 外文期刊>Journal of Molecular Biology >Dimerization of the 3'UTR of bicoid mRNA involves a two-step mechanism.
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Dimerization of the 3'UTR of bicoid mRNA involves a two-step mechanism.

机译：bicoid mRNA 3'UTR的二聚化涉及两步机制。

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摘要

The proper localization of bicoid (bcd) mRNA requires cis-acting signals within its 3' untranslated region (UTR) and trans-acting factors such as Staufen. Dimerization of bcd mRNA through intermolecular base-pairing between two complementary loops of domain III of the 3'UTR was proposed to be important for particle formation in the embryo. The participation in the dimerization process of each domain building the 3'UTR was evaluated by thermodynamic and kinetic analysis of various mutated and truncated RNAs. Although sequence complementarity between the two loops of domain III is required for initiating mRNA dimerization, the initial reversible loop-loop complex is converted rapidly into an almost irreversible complex. This conversion involves parts of RNA outside of domain III that promote initial recognition, and dimerization can be inhibited by sense or antisense oligonucleotides only before conversion has proceeded. Injection of the different bcd RNA variants into living Drosophila embryos shows that all elements that inhibit RNA dimerization in vitro prevent formation of localized particles containing Staufen. Particle formation appeared to be dependent on both mRNA dimerization and other element(s) in domains IV and V. Domain III of bcd mRNA could be substituted by heterologous dimerization motifs of different geometry. The resulting dimers were converted into stable forms, independently of the dimerization module used. Moreover, these chimeric RNAs were competent in forming localized particles and recruiting Staufen. The finding that the dimerization domain of bcd mRNA is interchangeable suggests that dimerization by itself, and not the precise geometry of the intermolecular interactions, is essential for the localization process. This suggests that the stabilizing interactions that are formed during the second step of the dimerization process might represent crucial elements for Staufen recognition and localization.
机译：bicoid（bcd）mRNA的正确定位需要其3'非翻译区（UTR）中的顺式作用信号和反式作用因子（例如Staufen）。通过在3'UTR结构域III的两个互补环之间的分子间碱基配对，使bcd mRNA的二聚化对于胚胎中颗粒的形成很重要。通过对各种突变和截短的RNA进行热力学和动力学分析，评估构建3'UTR的每个结构域的二聚化过程的参与。虽然域III的两个环之间的序列互补性对于启动mRNA二聚化是必需的，但最初的可逆环-环复合物会迅速转化为几乎不可逆的复合物。这种转化涉及结构域III之外的一部分RNA，这些RNA促进了最初的识别，并且只有在转化进行之前，才可以通过有义或反义寡核苷酸抑制二聚化。将不同的bcd RNA变体注入活的果蝇胚胎表明，所有在体外抑制RNA二聚化的元素都可以防止含有Staufen的局部颗粒的形成。颗粒的形成似乎既依赖于mRNA二聚化，也依赖于域IV和V中的其他元素。bcd mRNA的域III可以被不同几何结构的异源二聚化基序所取代。所得二聚体被转化为稳定形式，而与所使用的二聚模块无关。此外，这些嵌合RNA能够形成局部颗粒并募集Staufen。 bcd mRNA的二聚化结构域可互换的发现表明，二聚化本身而不是分子间相互作用的精确几何形状对于定位过程至关重要。这表明在二聚化过程的第二步过程中形成的稳定相互作用可能代表了Staufen识别和定位的关键要素。

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