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Translationally controlled tumor protein is required for the fast growth of Toxoplasma gondii and maintenance of its intracellular development
Author(s) -
Zheng Jun,
Chen Yaping,
Li Zhaoran,
Cao Shinuo,
Zhang Zhaoxia,
Jia Honglin
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.201700994r
Subject(s) - biology , gene , toxoplasma gondii , virulence , crispr , recombinant dna , genome , phenotype , genetics , microbiology and biotechnology , antibody
Translationally controlled tumor protein (TCTP) is a highly conserved, multifunctional protein that has been implicated in a range of cell physiologic processes, especially cell growth and development. A TCTP‐like gene has been identified in the Toxoplasma genome [ Toxoplasma gondii TCTP ( Tg TCTP)], although its function remains unknown. The sequence analysis of Tg TCTP indicated that it is a highly conserved protein in eukaryotes. We found that the expression level of Tg TCTP in the virulent RH strain was significantly higher than that in the avirulent PLK strain. Indirect immunofluorescence showed that Tg TCTP was expressed in the parasite cytoplasm. The localization of Tg TCTP was unchanged during the replication of the parasite. We expressed a functional recombinant Tg TCTP (r Tg TCTP) protein in Escherichia coli and found that the recombinant protein could form a multimer. We then evaluated the function of Tg TCTP using the clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR‐associated protein 9 knockout (KO) system. Phenotypic analysis of the KO strain (Δ Tg TCTP) revealed that Tg TCTP is required for the robust growth of the parasites. Tg TCTP deficiency also led to early egress of the parasites and subsequent impairment in their invasion and attachment abilities. We subsequently found that the multimer form of Tg TCTP might not be necessary for the growth and replication of the parasite. Then the expression profiling of genes in the Δ Tg TCTP and complement strains were analyzed. The results revealed that 988 genes were regulated in Δ Tg TCTP compared with the complement strain. Overall, although not essential, Tg TCTP is required for the fast growth of Tg and maintenance of its intracellular development.—Zheng J., Chen, Y., Li, Z., Cao, S., Zhang Z., Jia, H. Translationally controlled tumor protein is required for the fast growth of Toxoplasma gondii and maintenance of its intracellular development. FASEB J. 32, 906–919 (2018). www.fasebj.org