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Characterization of mitochondrion-targeted GTPases in Plasmodium falciparum

机译：疟原虫中线粒体靶向GTP酶的特征

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摘要

Ribosome assembly is critical for translation and regulating the response to cellular events and requires a complex interplay of ribosomal RNA and proteins with assembly factors. We investigated putative participants in the biogenesis of the reduced organellar ribosomes of Plasmodium falciparum and identified homologues of two assembly GTPases – EngA and Obg that were found in mitochondria. Both are indispensable in bacteria and P. berghei EngA is among the ‘essential’ parasite blood stage proteins identified recently. PfEngA and PfObg1 interacted with parasite mitoribosomes in vivo. GTP stimulated PfEngA interaction with the 50S subunit of Escherichia coli surrogate ribosomes. Although PfObg1–ribosome interaction was independent of nucleotide binding, GTP hydrolysis by PfObg1 was enhanced upon ribosomal association. An additional function for PfObg1 in mitochondrial DNA transactions was suggested by its specific interaction with the parasite mitochondrial genome in vivo. Deletion analysis revealed that the positively-charged OBG (spoOB-associated GTP-binding protein) domain mediates DNA-binding. A role for PfEngA in mitochondrial genotoxic stress response was indicated by its over-expression upon methyl methanesulfonate-induced DNA damage. PfEngA had lower sensitivity to an E. coli EngA inhibitor suggesting differences with bacterial counterparts. Our results show the involvement of two important GTPases in P. falciparum mitochondrial function, with the first confirmed localization of an EngA homologue in eukaryotic mitochondria.
机译：核糖体组件对于翻译和调节对细胞事件的响应至关重要，并且需要核糖体RNA和蛋白质的复杂相互作用。我们调查了在恶性疟原虫减少的细胞内核糖体的生物发生的推定参与者，并鉴定了在线粒体中发现的两个组装GTP酶的同源物的同源物。两者都是必不可少的细菌，P. Berghei Enga是最近发现的“必需的”寄生虫血液阶段蛋白质。 pfenga和pfobg1与体内寄生虫毛皮酶相互作用。 GTP刺激了与大肠杆菌替代核糖体的50s亚基的Pfenga相互作用。虽然pfobg1-核糖体相互作用与核苷酸结合无关，但在核糖体结合时通过pfobg1的GTP水解。通过其在体内寄生虫线粒体基因组的特定相互作用提出了线粒体DNA事务中PFOBG1的附加功能。缺失分析显示，带正电荷的OBG（纺织品相关的GTP结合蛋白）结构域介导DNA结合。通过其过表达对甲磺酸甲酯诱导的DNA损伤的过表达来表明pfenga在线粒体遗传毒性应激响应中的作用。 PFENGA对大肠杆菌ENGA抑制剂的敏感性较低，表明与细菌对应物的差异。我们的结果表明，两种重要的GTP酶在P. falciparum线粒体功能中的参与，并在真核线粒体中的首先确认了Enga同源物的定位。

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