Cells use fatty acids (FAs) for membrane biosynthesis, energy storage, and the generation of signaling molecules. 3-hydroxyacyl-CoA dehydratase-DEH-is a key component of very long chain fatty acid synthesis. Here, we further characterized in-depth the location and function of DEH, applying in silico analysis, live cell imaging, reverse genetics, and ultrastructure analysis using the mouse malaria model Plasmodium berghei DEH is evolutionarily conserved across eukaryotes, with a single DEH in Plasmodium spp. and up to three orthologs in the other eukaryotes studied. DEH-GFP live-cell imaging showed strong GFP fluorescence throughout the life-cycle, with areas of localized expression in the cytoplasm and a circular ring pattern around the nucleus that colocalized with ER markers. Δdeh mutants showed a small but significant reduction in oocyst size compared with WT controls from day 10 postinfection onwards, and endomitotic cell division and sporogony were completely ablated, blocking parasite transmission from mosquito to vertebrate host. Ultrastructure analysis confirmed degeneration of Δdeh oocysts, and a complete lack of sporozoite budding. Overall, DEH is evolutionarily conserved, localizes to the ER, and plays a crucial role in sporogony.

中文翻译：

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疟原虫 DEH 位于内质网，对于疟疾传播过程中卵囊有丝分裂至关重要。


细胞利用脂肪酸 (FA) 进行膜生物合成、能量储存和信号分子的生成。 3-羟酰基-CoA脱水酶-DEH-是极长链脂肪酸合成的关键成分。在这里，我们进一步深入表征了 DEH 的位置和功能，并应用于计算机分析、活细胞成像、反向遗传学和使用小鼠疟疾模型的超微结构分析。伯氏疟原虫DEH 在真核生物中进化上是保守的，疟原虫中只有一个 DEH种。在所研究的其他真核生物中最多有三个直系同源物。 DEH-GFP 活细胞成像在整个生命周期中显示出强烈的 GFP 荧光，在细胞质中具有局部表达区域，并且在细胞核周围有与 ER 标记共定位的圆环图案。从感染后第 10 天起，与 WT 对照相比，Δ deh突变体显示出卵囊大小小幅但显着的减少，并且内有丝分裂细胞分裂和孢子生殖完全消除，阻止了寄生虫从蚊子到脊椎动物宿主的传播。超微结构分析证实 Δ deh卵囊变性，并且完全缺乏子孢子出芽。总体而言，DEH 在进化上是保守的，定位于 ER，并且在孢子生殖中发挥着至关重要的作用。