Journal of Molecular Evolution

, Volume 60, Issue 1, pp 113–127 | Cite as

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

  • Takeshi Annoura
  • Takeshi Nara
  • Takashi Makiuchi
  • Tetsuo Hashimoto
  • Takashi Aoki


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.


Dihydroorotate dehydrogenase Trypanosoma cruzi Bodonid Kinetoplastid Euglenozoa Phylogenetic tree Gene fusion Horizontal gene transfer Anaerobiosis 



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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Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  • Takeshi Annoura
    • 1
  • Takeshi Nara
    • 1
  • Takashi Makiuchi
    • 1
    • 2
  • Tetsuo Hashimoto
    • 3
  • Takashi Aoki
    • 1
  1. 1.Department of Molecular and Cellular ParasitologyJuntendo University School of MedicineBunkyo-kuJapan
  2. 2.Department of Molecular BiologyTokyo University of Pharmacy and Life ScienceHachiojiJapan
  3. 3.Institute of Biological SciencesUniversity of TsukubaTsukubaJapan

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