Critical importance of the de novo pyrimidine biosynthesis pathway for Trypanosoma cruzi growth in the mammalian host cell cytoplasm

• Published in: Biochemical and biophysical research communications Vol. 417; no. 3; pp. 1002 - 1006
• Main Authors: Hashimoto, Muneaki, Morales, Jorge, Fukai, Yoshihisa, Suzuki, Shigeo, Takamiya, Shinzaburo, Tsubouchi, Akiko, Inoue, Syou, Inoue, Masayuki, Kita, Kiyoshi, Harada, Shigeharu, Tanaka, Akiko, Aoki, Takashi, Nara, Takeshi
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20.01.2012
• Subjects: 60 APPLIED LIFE SCIENCES; BIOSYNTHESIS; Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing) - genetics; Carbamoyl-phosphate synthetase II; Cell growth; CELL PROLIFERATION; Chagas Disease - metabolism; Chagas Disease - parasitology; CHEMOTHERAPY; CYTOPLASM; Cytoplasm - metabolism; Cytoplasm - parasitology; DISEASES; Gene Knockout Techniques; Gene targeting; GENES; HeLa Cells; Humans; INFUSION; INSECTS; LIGASES; LIVER; MUTANTS; NUCLEOTIDES; PHOSPHATES; Pyrimidine biosynthesis pathway; PYRIMIDINES; Pyrimidines - biosynthesis; TRYPANOSOMA; Trypanosoma cruzi; Trypanosoma cruzi - genetics; Trypanosoma cruzi - growth & development; TUMOR CELLS; Cell growth; CPSII; Gene targeting; LIT; MOI; Pyrimidine biosynthesis pathway; Carbamoyl-phosphate synthetase II; DHOD; Trypanosoma cruzi
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2011.12.073

► We established Trypanosoma cruzi lacking the gene for carbamoyl phosphate synthetase II. ► Disruption of the cpsII gene significantly reduced the growth of epimastigotes. ► In particular, the CPSII-null mutant severely retarded intracellular growth. ► The de novo pyrimidine pathway is critical for the parasite growth in the host cell. The intracellular parasitic protist Trypanosoma cruzi is the causative agent of Chagas disease in Latin America. In general, pyrimidine nucleotides are supplied by both de novo biosynthesis and salvage pathways. While epimastigotes—an insect form—possess both activities, amastigotes—an intracellular replicating form of T. cruzi—are unable to mediate the uptake of pyrimidine. However, the requirement of de novo pyrimidine biosynthesis for parasite growth and survival has not yet been elucidated. Carbamoyl-phosphate synthetase II (CPSII) is the first and rate-limiting enzyme of the de novo biosynthetic pathway, and increased CPSII activity is associated with the rapid proliferation of tumor cells. In the present study, we showed that disruption of the T. cruzicpsII gene significantly reduced parasite growth. In particular, the growth of amastigotes lacking the cpsII gene was severely suppressed. Thus, the de novo pyrimidine pathway is important for proliferation of T. cruzi in the host cell cytoplasm and represents a promising target for chemotherapy against Chagas disease.