Inducing controlled cell cycle arrest and re-entry during asexual proliferation of Plasmodium falciparum malaria parasites.
Inducing controlled cell cycle arrest and re-entry during asexual proliferation of Plasmodium falciparum malaria parasites.
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DOI：
10.1038/s41598-018-34964-w
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发表时间：
2018-11-08
影响因子：
4.6
通讯作者：
Birkholtz LM
中科院分区：
文献类型：
--
作者：
van Biljon R;Niemand J;van Wyk R;Clark K;Verlinden B;Abrie C;von Grüning H;Smidt W;Smit A;Reader J;Painter H;Llinás M;Doerig C;Birkholtz LM
疟疾寄生虫恶性疟原虫的生命周期受到严格调控，在剧烈增殖和静止阶段之间摇摆。疟原虫的48小时周期性无性复制与高等真核生物的细胞分裂明显不同，其机制尚不清楚。在这里，我们报告了在细胞周期的早期阶段，通过暴露于DL-α-二氟甲基鸟氨酸（DFMO），这导致多胺的耗尽疟疾寄生虫的紧密同步。这通过阻断G1/S转换而诱导不可避免的细胞周期停滞在G1（侵入后约15小时）。细胞周期停滞的寄生虫进入静止的G 0样状态，但在加入外源性多胺后，重新启动其细胞周期。这种将疟原虫停止在其细胞周期中的特定点并随后触发重新进入细胞周期的能力为研究这些寄生虫的细胞周期调控提供了有价值的框架。随后，我们使用基因表达分析表明，重新进入细胞周期涉及表达钙敏感（cdpk 4和pk 2）和有丝分裂激酶（nima和ark 2），与失调的复制前复合物与pk 2的表达。基因表达的变化可以通过转录因子MYB 1和两个ApiAP 2家族成员驱动。因此，这种寄生虫同步的新方法扩展了我们目前有限的工具包，以研究疟疾寄生虫的细胞周期调控。
The life cycle of the malaria parasite Plasmodium falciparum is tightly regulated, oscillating between stages of intense proliferation and quiescence. Cyclic 48-hour asexual replication of Plasmodium is markedly different from cell division in higher eukaryotes, and mechanistically poorly understood. Here, we report tight synchronisation of malaria parasites during the early phases of the cell cycle by exposure to DL-α-difluoromethylornithine (DFMO), which results in the depletion of polyamines. This induces an inescapable cell cycle arrest in G1 (~15 hours post-invasion) by blocking G1/S transition. Cell cycle-arrested parasites enter a quiescent G0-like state but, upon addition of exogenous polyamines, re-initiate their cell cycle. This ability to halt malaria parasites at a specific point in their cell cycle, and to subsequently trigger re-entry into the cell cycle, provides a valuable framework to investigate cell cycle regulation in these parasites. We subsequently used gene expression analyses to show that re-entry into the cell cycle involves expression of Ca2+-sensitive (cdpk4 and pk2) and mitotic kinases (nima and ark2), with deregulation of the pre-replicative complex associated with expression of pk2. Changes in gene expression could be driven through transcription factors MYB1 and two ApiAP2 family members. This new approach to parasite synchronisation therefore expands our currently limited toolkit to investigate cell cycle regulation in malaria parasites.
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影响因子：
12.3
作者：
Aragon AD;Quiñones GA;Thomas EV;Roy S;Werner-Washburne M
通讯作者：
Werner-Washburne M
激光捕获显微解剖可以实现疟原虫复发物种的分裂和静止肝阶段的转录组分析。
影响因子：
3.4
作者：
Cubi R;Vembar SS;Biton A;Franetich JF;Bordessoulles M;Sossau D;Zanghi G;Bosson-Vanga H;Benard M;Moreno A;Dereuddre-Bosquet N;Le Grand R;Scherf A;Mazier D
通讯作者：
Mazier D
影响因子：
9.8
作者：
Bozdech, Zbynek;Llinas, Manuel;Pulliam, Brian Lee;Wong, Edith D;Zhu, Jingchun;DeRisi, Joseph L
通讯作者：
DeRisi, Joseph L
影响因子：
3.6
作者：
Deshmukh, Abhijit S.;Agarwal, Meetu;Dhar, Suman Kumar
通讯作者：
Dhar, Suman Kumar
影响因子：
4.9
作者：
Das Gupta, R;Krause-Ihle, T;Lüersen, K
通讯作者：
Lüersen, K