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The Origin of Dihydroorotate Dehydrogenase Genes of Kinetoplastids, with Special Reference to Their Biological Significance and Adaptation to Anaerobic, Parasitic Conditions

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Abstract

Trypanosoma cruzi dihydroorotate dehydrogenase (DHOD), the fourth enzyme of the de novo pyrimidine biosynthetic pathway, is localized in the cytosol and utilizes fumarate as electron acceptor (fumarate reductase activity), while the enzyme from other various eukaryotes is mitochondrial membrane-linked. Here we report that DHOD-knockout T. cruzi did not express the enzyme protein and could not survive even in the presence of pyrimidine nucleosides, substrates for the potentially active salvage pathway, suggesting a vital role of fumarate reductase activity in the regulation of cellular redox balance. Cloning and phylogenetic analysis of euglenozoan DHOD genes showed that the euglenoid Euglena gracilis had a mitochondrial DHOD and that biflagellated bodonids, a sister group of trypanosomatids within kinetoplastids, harbor the cytosolic DHOD. Further, Bodo saliens, a bodonid, had an ACT/DHOD gene fusion encoding aspartate carbamoyltransferase (ACT), the second enzyme of the de novo pyrimidine pathway, and DHOD. This is the first report of this novel gene structure. These results are consistent with suggestions that an ancient common ancestor of Euglenozoa had a mitochondrial DHOD whose descendant exists in E. gracilis and that a common ancestor of kinetoplastids (bodonids and trypanosomatids) subsequently acquired a cytosolic DHOD by horizontal gene transfer. The cytosolic DHOD gene thus acquired may have contributed to adaptation to anaerobiosis in the kinetoplastid lineage and further contributed to the subsequent establishment of parasitism in a trypanosomatid ancestor. Different molecular strategies for anaerobic adaptation in pyrimidine biosynthesis, used by kinetoplastids and by euglenoids, are discussed. Evolutionary implications of the ACT/DHOD gene fusion are also discussed.

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Acknowledgments

We express our sincere thanks to Dr. Miklós Müller for his critical reading of the manuscript, Mr. Masao Odaka and Ms. Yuko Mikami for their technical assistance, and Dr. Tomoyoshi Nozaki for plasmids p72hyg72 and p72tun72. A part of this work was supported by grants-in-aid for scientific research (Nos. 15390138, 15659102, and 13640709) from the Ministry of Education, Science, Sports, and Culture of Japan, from Ohyama Health Foundation (to T.N.), and from Kampou Science Foundation (to T.N.). T. Annoura and T. Aoki are supported by a Grant-in-Aid for 21st Century COE Research from the Ministry of Education, Science, Sports, and Culture of Japan.

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Authors and Affiliations

  1. Department of Molecular and Cellular Parasitology, Juntendo University School of Medicine, Hongo 2-1-1, Bunkyo-ku, Tokyo, 113-8421, Japan

    Takeshi Annoura, Takeshi Nara, Takashi Makiuchi & Takashi Aoki

  2. Department of Molecular Biology, Tokyo University of Pharmacy and Life Science, Horinouchi 1432-1, Hachioji, Tokyo, 192-0392, Japan

    Takashi Makiuchi

  3. Institute of Biological Sciences, University of Tsukuba, Tennoudai 1-1-1, Tsukuba, Ibaraki, 305-8575, Japan

    Tetsuo Hashimoto

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  1. Takeshi Annoura
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  2. Takeshi Nara
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  4. Tetsuo Hashimoto
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  5. Takashi Aoki
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Correspondence to Takeshi Nara.

Additional information

Sequence availability: The nucleotide sequence data reported here appear in the GenBank, EMBL, and DDBJ databases with the accession numbers AB120414, AB159227, and AB159228 for Euglena gracilis dihydroorotate dehydrogenase (DHOD), Bodo saliens aspartate carbamoyltransferase/dihydroorotate dehydrogenase (ACT/DHOD), and B. caudatus DHOD, respectively.

Reviewing Editor: Dr. Patrick Keeling

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Annoura, T., Nara, T., Makiuchi, T. et al. The Origin of Dihydroorotate Dehydrogenase Genes of Kinetoplastids, with Special Reference to Their Biological Significance and Adaptation to Anaerobic, Parasitic Conditions. J Mol Evol 60, 113–127 (2005). https://doi.org/10.1007/s00239-004-0078-8

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