Plasmodium falciparum Calcium-Dependent Protein Kinase 4 is Critical for Male Gametogenesis and Transmission to the Mosquito Vector.
Plasmodium falciparum Calcium-Dependent Protein Kinase 4 is Critical for Male Gametogenesis and Transmission to the Mosquito Vector.
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DOI：
10.1128/mbio.02575-21
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发表时间：
2021-12-21
期刊：
影响因子：
6.4
通讯作者：
Swearingen KE
中科院分区：
文献类型：
--
作者：
Kumar S;Haile MT;Hoopmann MR;Tran LT;Michaels SA;Morrone SR;Ojo KK;Reynolds LM;Kusebauch U;Vaughan AM;Moritz RL;Kappe SHI;Swearingen KE
疟疾寄生虫疟原虫的配子体被蚊子媒介以传染性血液为食，这是寄生虫传播的关键阶段。钙非依赖性蛋白激酶（CDPKs）在寄生虫复杂的生命周期中钙介导的信号传导中发挥关键作用。我们试图了解它们在人类寄生虫从宿主传播到蚊子载体中的作用，从而研究人类感染的恶性疟原虫CDPK4在寄生虫生命周期中的作用。靶基因缺失产生的恶性疟原虫cdpk4 -寄生虫对血期发育或配子体发育没有影响。然而，cdpk4−寄生虫在雄性配子发生和鞭毛雄性配子的出现方面存在严重缺陷。为了了解这种缺陷的分子基础，我们对野生型和恶性疟原虫cdpk4 -晚期配子细胞阶段进行了基于质谱的磷酸化蛋白质组学分析，以确定cdpk4介导的关键磷酸化事件，这些磷酸化事件可能对雄性配子发生的调节很重要。我们进一步采用体外实验来鉴定这些假定的恶性疟原虫CDPK4底物。这表明CDPK4通过直接或间接控制DNA复制、mRNA翻译和细胞运动等关键事件来调节雄性配子体发生。综上所述，我们的工作表明PfCDPK4是一种调节鞭毛的中心激酶，因此对寄生虫传播到蚊子载体至关重要。
Gametocytes of the malaria parasite Plasmodium are taken up by the mosquito vector with an infectious blood meal, representing a critical stage for parasite transmission. Calcium-independent protein kinases (CDPKs) play key roles in calcium-mediated signaling across the complex life cycle of the parasite. We sought to understand their role in human parasite transmission from the host to the mosquito vector and thus investigated the role of the human-infective parasite Plasmodium falciparum CDPK4 in the parasite life cycle. P. falciparum cdpk4− parasites created by targeted gene deletion showed no effect in blood stage development or gametocyte development. However, cdpk4− parasites showed a severe defect in male gametogenesis and the emergence of flagellated male gametes. To understand the molecular underpinnings of this defect, we performed mass spectrometry-based phosphoproteomic analyses of wild-type and Plasmodium falciparum cdpk4− late gametocyte stages to identify key CDPK4-mediated phosphorylation events that may be important for the regulation of male gametogenesis. We further employed in vitro assays to identify these putative substrates of Plasmodium falciparum CDPK4. This indicated that CDPK4 regulates male gametogenesis by directly or indirectly controlling key essential events, such as DNA replication, mRNA translation, and cell motility. Taken together, our work demonstrates that PfCDPK4 is a central kinase that regulates exflagellation and thereby is critical for parasite transmission to the mosquito vector.