Pb103 Regulates Zygote/Ookinete Development in Plasmodium berghei via Double Zinc Finger Domains.
Pb103 Regulates Zygote/Ookinete Development in Plasmodium berghei via Double Zinc Finger Domains.
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DOI：
10.3390/pathogens10121536
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发表时间：
2021-11-24
期刊：
影响因子：
--
通讯作者：
Mita T
中科院分区：
文献类型：
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作者：
Hirai M;Maeta A;Mori T;Mita T
疟原虫寄生虫的有性繁殖发生在按蚊中，雄性和雌性配子融合形成受精卵，然后形成动合子。这些过程是由阶段特异性蛋白质表达精心策划的，而这部分是由翻译抑制介导的。越来越多的证据表明 RNA 结合蛋白 (RBP) 在这些过程中发挥着至关重要的作用。在这里，我们报告了 P. berghei 103 (Pb103) 的特征，它编码具有双锌指结构域 (ZF) 的蛋白质，即 RBP。在内源性 Pb103 基因启动子（Pb103-AGFP 报告基因）下表达 azami 绿色荧光蛋白（AGFP）的报告寄生虫表明，从配子到动合子都能检测到 AGFP 荧光信号，而雌配子细胞中 AGFP mRNA 的翻译受到抑制。 Pb103 破坏的寄生虫 (Pb103(−)) 生长并产生配子体，其效率与野生型寄生虫相似。然而，喂食 Pb103(−) 的蚊子没有形成卵囊。体外受精试验显示 Pb103(−) 在受精卵阶段发生流产，表明 Pb103 在受精卵/动合子发育中发挥着关键作用。 Pb103(−) 与雄性或雌性不育寄生虫的杂交受精测定表明，Pb103 仅对雌性配子至关重要。为了确定对合子/动动体发育至关重要的结构域，生成了表达部分缺失的 Pb103 的转基因寄生虫并分析了动动体的成熟。因此，删除两个 ZF 中的任何一个（但不删除 C 末端区域）会废除合子/动合子的发育，突出了 ZF 在寄生虫性发育中不可或缺的作用，很可能是通过翻译抑制。
Sexual reproduction of Plasmodium parasites takes place in anopheline mosquitoes, where male and female gametes fuse to form zygotes and then ookinetes. These processes are orchestrated by stage-specific protein expression, which is mediated in part by translational repression. Accumulating evidence shows that RNA binding proteins (RBPs) play crucial roles in these processes. Here, we report the characterization of P. berghei 103 (Pb103), which encodes a protein possessing double zinc finger domains (ZFs), an RBP. Reporter parasites expressing azami green fluorescent protein (AGFP) under the endogenous Pb103 gene promoter (Pb103-AGFP reporter) showed that the AGFP fluorescent signal was detected from gametes to ookinetes, while AGFP mRNA was translationally repressed in female gametocytes. The Pb103-disrupted parasites (Pb103(−)) grew and produced gametocytes with similar efficiencies to those of wild-type parasites. However, no oocysts were formed in mosquitoes fed Pb103(−). An in vitro fertilization assay showed abortion at the zygote stage in Pb103(−), suggesting that Pb103 plays a critical role in zygote/ookinete development. Cross-fertilization assays with Pb103(−) and male- or female-sterile parasites revealed that Pb103 was essential exclusively for female gametes. To identify the domains critical for zygote/ookinete development, transgenic parasites expressing partially deleted Pb103 were generated and assayed for ookinete maturation. As a result, deleting either of two ZFs but not the C-terminal region abolished zygote/ookinete development, highlighting the indispensable roles of ZFs in parasite sexual development, most likely via translational repression.
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