cAMP-dependent protein kinase regulates secretion of apical membrane antigen 1 (AMA1) in Plasmodium yoelii
cAMP-dependent protein kinase regulates secretion of apical membrane antigen 1 (AMA1) in Plasmodium yoelii
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cAMP依赖性蛋白激酶调节约氏疟原虫顶膜抗原1（AMA1）的分泌
DOI：
10.1016/j.parint.2021.102435
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发表时间：
2021
影响因子：
1.9
通讯作者：
Osamu Kaneko
中科院分区：
文献类型：
--
作者：
Takahiro Ishizaki;Masahito Asada;Hassan Hakimi;Nattawat Chaiyawong;Yuto Kegawa;Kazuhide Yahatab;Osamu Kaneko
疟疾仍然是全球人类健康的沉重负担，了解寄生虫的分子和细胞生物学以找到药物和疫苗开发的目标非常重要。小鼠疟疾模型是表征已鉴定分子功能的重要工具;然而，针对广泛使用的啮齿动物疟疾寄生虫约氏疟原虫的靶向基因缺失的稳健技术仍然开发不足。为了克服这一问题，我们建立了一个DiCre-loxP诱导敲除（iKO）系统在P。yoelii，其显示出在RAP施用后24小时（一个细胞周期）超过80%的靶基因座切除效率和超过90%的基因座转录物减少。使用这种开发的系统，诱导破坏cAMP依赖性蛋白激酶（PKAc），并分析所得PKAc-iKO寄生虫的表型。我们发现PKAc-iKO寄生虫表现出严重的生长和红细胞侵袭缺陷。我们还发现，PKAc的破坏损害了AMA 1在P中的分泌。yoelii，与一份显示PKAc在P中AMA 1分泌中没有作用的报告相反。恶性疟原虫。这种差异可能与AMA 1分布到裂殖子表面的时间差异有关，裂殖子表面发生在P出口之后。恶性疟原虫，但几分钟后为P. yoelii。PyEBL、Py 235和RON 2的分泌不受PKAc在P中的破坏的影响。yoelii。PyRON 2在裂殖子排出后立即分泌到裂殖子表面，这与RON 2被注射到红细胞胞质溶胶中的当前模型不一致。需要进一步的研究来了解暴露在裂殖子表面的RON 2的作用。
Malaria remains a heavy global burden on human health, and it is important to understand the molecular and cellular biology of the parasite to find targets for drug and vaccine development. The mouse malaria model is an essential tool to characterize the function of identified molecules; however, robust technologies for targeted gene deletions are still poorly developed for the widely used rodent malaria parasite,Plasmodium yoelii. To overcome this problem, we established a DiCre-loxP inducible knockout (iKO) system inP. yoelii, which showed more than 80% excision efficacy of the target locus and more than 90% reduction of locus transcripts 24 h (one cell cycle) after RAP administration. Using this developed system, cAMP-dependent protein kinase (PKAc) was inducibly disrupted and the phenotypes of the resulting PKAc-iKO parasites were analyzed. We found that PKAc-iKO parasites showed severe growth and erythrocyte invasion defects. We also found that disruption of PKAc impaired the secretion of AMA1 inP. yoelii, in contrast to a report showing no role of PKAc in AMA1 secretion inP. falciparum. This discrepancy may be related to the difference in the timing of AMA1 distribution to the merozoite surface, which occurs just after egress forP. falciparum, but after several minutes forP. yoelii. Secretions of PyEBL, Py235, and RON2 were not affected by the disruption of PKAc inP. yoelii. PyRON2 was already secreted to the merozoite surface immediately after merozoite egress, which is inconsistent with the current model that RON2 is injected into the erythrocyte cytosol. Further investigations are required to understand the role of RON2 exposed on the merozoite surface.