Translationally controlled tumor protein is required for the fast growth of Toxoplasma gondii and maintenance of its intracellular development
Translationally controlled tumor protein is required for the fast growth of Toxoplasma gondii and maintenance of its intracellular development
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翻译控制的肿瘤蛋白是弓形虫快速生长和维持其细胞内发育所必需的
DOI：
10.1096/fj.201700994r
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发表时间：
2018-02-01
期刊：
影响因子：
4.8
通讯作者：
Jia, Honglin
中科院分区：
文献类型：
--
作者：
Zheng, Jun;Chen, Yaping;Jia, Honglin
翻译控制肿瘤蛋白（Translationally controlled tumor protein，TCTP）是一种高度保守的多功能蛋白质，参与细胞的生长发育过程。在弓形虫基因组中发现了ATCTP样基因[弓形虫TCTP（Toxoplasma gondii TCTP，TgTCTP）]，但其功能尚不清楚。序列分析表明，TgTCTP在真核生物中是一个高度保守的蛋白。我们发现，TgTCTP在强毒RH株中的表达水平显著高于无毒PLK株。间接免疫荧光显示TgTCTP在疟原虫胞质中表达。TgTCTP的定位在寄生虫的复制过程中是不变的。我们在大肠杆菌中表达了功能性重组TgTCTP（rTgTCTP）蛋白，并发现重组蛋白可以形成多聚体。然后，我们使用成簇规则间隔短回文重复序列（CRISPR）/CRISPR相关蛋白9敲除（KO）系统评估了TgTCTP的功能。KO菌株（Delta TgTCTP）的表型分析显示，TgTCTP是寄生虫稳健生长所必需的。TgTCTP缺陷也导致早期出口的寄生虫和随后的损害，在他们的入侵和附着能力。我们随后发现，TgTCTP的多聚体形式可能不是必要的寄生虫的生长和复制。然后分析Delta TgTCTP和补体菌株中基因的表达谱。结果显示，与补体菌株相比，Δ TgTCTP中有988个基因受到调控。总体而言，虽然不是必需的，但TgTCTP是Tg快速生长和维持其细胞内发育所必需的。
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. ATCTP-like gene has been identified in the Toxoplasma genome [Toxoplasma gondii TCTP (TgTCTP)], although its function remains unknown. The sequence analysis of TgTCTP indicated that it is a highly conserved protein in eukaryotes. We found that the expression level of TgTCTP in the virulent RH strain was significantly higher than that in the avirulent PLK strain. Indirect immunofluorescence showed that TgTCTP was expressed in the parasite cytoplasm. The localization of TgTCTP was unchanged during the replication of the parasite. We expressed a functional recombinant TgTCTP (rTgTCTP) protein in Escherichia coli and found that the recombinant protein could form a multimer. We then evaluated the function of TgTCTP using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 knockout (KO) system. Phenotypic analysis of the KO strain (Delta TgTCTP) revealed that TgTCTP is required for the robust growth of the parasites. TgTCTP deficiency also led to early egress of the parasites and subsequent impairment in their invasion and attachment abilities. We subsequently found that the multimer formof TgTCTP might not be necessary for the growth and replication of the parasite. Then the expression profiling of genes in the Delta TgTCTP and complement strains were analyzed. The results revealed that 988 genes were regulated in Delta TgTCTP compared with the complement strain. Overall, although not essential, TgTCTP is required for the fast growth of Tg and maintenance of its intracellular development.