Functional genomics of RAP proteins and their role in mitoribosome regulation in Plasmodium falciparum.
Functional genomics of RAP proteins and their role in mitoribosome regulation in Plasmodium falciparum.
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DOI：
10.1038/s41467-022-28981-7
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发表时间：
2022-03-11
影响因子：
16.6
通讯作者：
Le Roch KG
中科院分区：
文献类型：
--
作者：
Hollin T;Abel S;Falla A;Pasaje CFA;Bhatia A;Hur M;Kirkwood JS;Saraf A;Prudhomme J;De Souza A;Florens L;Niles JC;Le Roch KG
RAP(RNA结合结构域在复合体中丰富)蛋白家族已在多种生物中被鉴定。尽管该蛋白家族在顶端复合体寄生虫中扩大，但其主要生物学功能仍不清楚。在这项研究中，我们利用恶性疟原虫的诱导敲除研究，证明了两个RAP蛋白，PF3D7_0105200(PfRAP01)和PF3D7_1470600(PfRAP21)是寄生虫生存所必需的，并定位于线粒体。通过转录组学、代谢组学和蛋白质组学实验，我们进一步证明这些RAP蛋白参与了线粒体RNA的代谢。使用高通量测序的交联式免疫沉淀分离的RNA(eCLIP-seq)，我们验证了PfRAP01和PfRAP21是真正的RNA结合蛋白，并与线粒体rRNA特异地相互作用。最后，线粒体浓缩实验和小RNA的深度测序表明，PfRAP21控制着线粒体rRNA的表达。总之，我们的结果确定了这些RAP蛋白在有丝分裂体活性中的作用，并有助于进一步了解疟疾寄生虫中的这个蛋白质家族。RAP蛋白家族成员丰富的RNA结合域的功能在很大程度上是未知的。在这里，使用高通量功能基因组学，包括代谢组学，Hollin等人。鉴定恶性疟原虫生存和控制线粒体rRNAs所必需的两种RAP蛋白。
The RAP (RNA-binding domain abundant in Apicomplexans) protein family has been identified in various organisms. Despite expansion of this protein family in apicomplexan parasites, their main biological functions remain unknown. In this study, we use inducible knockdown studies in the human malaria parasite, Plasmodium falciparum, to show that two RAP proteins, PF3D7_0105200 (PfRAP01) and PF3D7_1470600 (PfRAP21), are essential for parasite survival and localize to the mitochondrion. Using transcriptomics, metabolomics, and proteomics profiling experiments, we further demonstrate that these RAP proteins are involved in mitochondrial RNA metabolism. Using high-throughput sequencing of RNA isolated by crosslinking immunoprecipitation (eCLIP-seq), we validate that PfRAP01 and PfRAP21 are true RNA-binding proteins and interact specifically with mitochondrial rRNAs. Finally, mitochondrial enrichment experiments followed by deep sequencing of small RNAs demonstrate that PfRAP21 controls mitochondrial rRNA expression. Collectively, our results establish the role of these RAP proteins in mitoribosome activity and contribute to further understanding this protein family in malaria parasites. The function of RNA-binding domain abundant in Apicomplexans (RAP) protein family members is largely unknown. Here, using high-throughput functional genomics, including metabolomics, Hollin et al. characterize two RAP proteins that are essential for Plasmodium falciparum survival and control mitochondrial rRNAs.
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影响因子：
14.9
作者：
Deitsch, KW;Driskill, CL;Wellems, TE
通讯作者：
Wellems, TE
影响因子：
7.4
作者：
Broeckling, C. D.;Afsar, F. A.;Prenni, J. E.
通讯作者：
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影响因子：
14.9
作者：
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通讯作者：
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DOI：
10.1073/pnas.1919501117
发表时间：
2020-06-16
影响因子：
11.1
作者：
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通讯作者：
Muralidharan, Vasant
影响因子：
16.6
作者：
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通讯作者：
Niles, Jacquin C.