Biochemical and functional analysis of Drosophila-sciara chimeric sex-lethal proteins.
Biochemical and functional analysis of Drosophila-sciara chimeric sex-lethal proteins.
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DOI：
10.1371/journal.pone.0065171
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Sánchez L
中科院分区：
文献类型：
--
作者：
Ruiz MF;Sarno F;Zorrilla S;Rivas G;Sánchez L
果蝇SXL蛋白控制性别决定和剂量补偿。它是一个性别特异的因子，控制着自身SXL Pre-mRNA的剪接(自动调节)、tra Pre-mRNA的剪接(性别决定)和msl-2 Pre-mRNA的翻译(剂量补偿)。在果蝇外，在两性中都发现了相同的SXL蛋白，因此，在非果蝇中，SXL似乎不像在果蝇中那样在性别决定和剂量补偿方面发挥关键的区分作用。SXL蛋白的比较表明，其空间结构是保守的，RNA结合区高度保守，而N-末端和C-末端结构域显示出显著的变异。这篇手稿关注的是SXL蛋白本身的进化，而不是它的表达调控。获得了果蝇-Sciara嵌合SXL蛋白。Sciara SXL代表了古代SXL在非果蝇中的非性别特异性功能，推测果蝇SXL是从这些非果蝇进化而来的。回答了两个问题。当果蝇SXL蛋白的N-末端和C-末端结构域被相应的SCHARA结构域取代时，SXL蛋白是否影响了它们的功能？当果蝇的N-末端和C-末端结构域取代Sciara的N-末端和C-末端结构域时，Sciara SXL蛋白是否获得了果蝇的性别特异性功能？体外分析嵌合的SXL蛋白以研究其结合亲和力和协同特性，并通过产生转SXL嵌合蛋白的果蝇体内分析其对性别决定和剂量补偿的影响。现存的果蝇SXL蛋白的性别特异性取决于其整体结构，而不是特定的结构域。这意味着SXL蛋白在果蝇谱系中进化过程中发生的修饰，主要发生在N-末端和C-末端区域，是共同进化的变化，决定了SXL执行其性别特异性功能的适当折叠。
The Drosophila SXL protein controls sex determination and dosage compensation. It is a sex-specific factor controlling splicing of its own Sxl pre-mRNA (auto-regulation), tra pre-mRNA (sex determination) and msl-2 pre-mRNA plus translation of msl-2 mRNA (dosage compensation). Outside the drosophilids, the same SXL protein has been found in both sexes so that, in the non-drosophilids, SXL does not appear to play the key discriminating role in sex determination and dosage compensation that it plays in Drosophila. Comparison of SXL proteins revealed that its spatial organisation is conserved, with the RNA-binding domains being highly conserved, whereas the N- and C-terminal domains showing significant variation. This manuscript focuses on the evolution of the SXL protein itself and not on regulation of its expression. Drosophila-Sciara chimeric SXL proteins were produced. Sciara SXL represents the non-sex-specific function of ancient SXL in the non-drosophilids from which presumably Drosophila SXL evolved. Two questions were addressed. Did the Drosophila SXL protein have affected their functions when their N- and C-terminal domains were replaced by the corresponding ones of Sciara? Did the Sciara SXL protein acquire Drosophila sex-specific functions when the Drosophila N- and C-terminal domains replaced those of Sciara? The chimeric SXL proteins were analysed in vitro to study their binding affinity and cooperative properties, and in vivo to analyse their effect on sex determination and dosage compensation by producing Drosophila flies that were transgenic for the chimeric SXL proteins. The sex-specific properties of extant Drosophila SXL protein depend on its global structure rather than on a specific domain. This implies that the modifications, mainly in the N- and C-terminal domains, that occurred in the SXL protein during its evolution within the drosophilid lineage represent co-evolutionary changes that determine the appropriate folding of SXL to carry out its sex-specific functions.
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