Mailu Boniface M, Li Ling, Arthur Jen, Nelson Todd M, Ramasamy Gowthaman, Fritz-Wolf Karin, Becker Katja, Gardner Malcolm J
From the Center for Infectious Disease Research, Seattle, Washington 98109.
the Department of Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University, Giessen 35392 Germany, and the Max-Planck Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany.
J Biol Chem. 2015 Dec 4;290(49):29629-41. doi: 10.1074/jbc.M115.655100. Epub 2015 Aug 28.
The malaria parasite Plasmodium falciparum apicoplast indirect aminoacylation pathway utilizes a non-discriminating glutamyl-tRNA synthetase to synthesize Glu-tRNA(Gln) and a glutaminyl-tRNA amidotransferase to convert Glu-tRNA(Gln) to Gln-tRNA(Gln). Here, we show that Plasmodium falciparum and other apicomplexans possess a unique heterodimeric glutamyl-tRNA amidotransferase consisting of GatA and GatB subunits (GatAB). We localized the P. falciparum GatA and GatB subunits to the apicoplast in blood stage parasites and demonstrated that recombinant GatAB converts Glu-tRNA(Gln) to Gln-tRNA(Gln) in vitro. We demonstrate that the apicoplast GatAB-catalyzed reaction is essential to the parasite blood stages because we could not delete the Plasmodium berghei gene encoding GatA in blood stage parasites in vivo. A phylogenetic analysis placed the split between Plasmodium GatB, archaeal GatE, and bacterial GatB prior to the phylogenetic divide between bacteria and archaea. Moreover, Plasmodium GatA also appears to have emerged prior to the bacterial-archaeal phylogenetic divide. Thus, although GatAB is found in Plasmodium, it emerged prior to the phylogenetic separation of archaea and bacteria.
恶性疟原虫的质体间接氨酰化途径利用一种无特异性的谷氨酰胺-tRNA合成酶来合成Glu-tRNA(Gln),并利用谷氨酰胺-tRNA氨基转移酶将Glu-tRNA(Gln)转化为Gln-tRNA(Gln)。在此,我们发现恶性疟原虫和其他顶复门原虫拥有一种独特的异源二聚体谷氨酰胺-tRNA氨基转移酶,它由GatA和GatB亚基(GatAB)组成。我们将恶性疟原虫的GatA和GatB亚基定位于血液期寄生虫的质体中,并证明重组GatAB在体外可将Glu-tRNA(Gln)转化为Gln-tRNA(Gln)。我们证明质体GatAB催化的反应对寄生虫血液期至关重要,因为我们无法在体内删除伯氏疟原虫中编码GatA的基因。系统发育分析表明,疟原虫GatB、古菌GatE和细菌GatB之间的分化发生在细菌和古菌的系统发育分化之前。此外,疟原虫GatA似乎也在细菌-古菌系统发育分化之前就已出现。因此,虽然GatAB存在于疟原虫中,但它在古菌和细菌的系统发育分离之前就已出现。