Characterization of a novel Obg-like ATPase in the protozoan Trypanosoma cruzi
Characterization of a novel Obg-like ATPase in the protozoan Trypanosoma cruzi
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DOI：
10.1016/j.ijpara.2008.05.019
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发表时间：
2009-01-01
影响因子：
4
通讯作者：
Fragoso, Stenio P.
中科院分区：
文献类型：
--
作者：
Gradia, Daniela F.;Rau, Karlan;Fragoso, Stenio P.
我们表征了编码YchF相关蛋白TcYchF的基因，该基因可能与克氏锥虫的蛋白质翻译机制相关。YchF属于翻译因子相关（TRAFAC）类的P-环NTPases。该基因的编码区长1185 bp，编码44.3 kDa的蛋白质。BlastX搜索显示TcYchF与真核生物中推定的GP结合蛋白非常相似（45-86%），包括一些锥虫（利什曼原虫和布氏锥虫）。在SpoOB相关GTP结合蛋白（Obg）家族的TcYchF和细菌[YyaF/YchF GTP酶]的预测序列之间也发现了较低但显著水平的相似性（38-43%）。这个GTP酶家族的G结构域的一些最重要的特征在TcYchF中是保守的。然而，我们发现TcYchF优先水解ATP而不是GTP。YyaF/YchF的功能是，但已知Obg家族的其他成员与核糖体亚基相关。未知的，来自蔗糖梯度的多核糖体组分的免疫印迹显示TcYchF与核糖体亚基和多核糖体相关。免疫沉淀实验表明，TcYchF还与T.克鲁兹此外，T. TcYchF的布氏同源物通过RNA干扰抑制寄生虫的前环形式的生长。这些数据表明，这种蛋白质在锥虫的翻译机制中起着重要作用。(C)2008年澳大利亚寄生虫学会由爱思唯尔有限公司出版。保留所有权利。
We characterized a gene encoding an YchF-related protein, TcYchF, potentially associated with the protein translation machinery of Trypanosoma cruzi. YchF belongs to the translation factor-related (TRAFAC) class of P-loop NTPases. The coding region of the gene is 1185 bp long and encodes a 44.3 kDa protein. BlastX searches showed TcYchF to be very similar (45-86%) to putative GTP-binding proteins from eukaryotes, including some species of trypanosomatids (Leishmania major and Trypanosoma brucei). A lower but significant level of similarity (38-43%) was also found between the predicted sequences of TcYchF and bacteria] YyaF/YchF GTPases of the SpoOB-associated GTP-binding protein (Obg) family. Some of the most important features of the G domain of this family of GTPases are conserved in TcYchF. However, we found that TcYchF preferentially hydrolyzed ATP rather than GTP. The function of YyaF/YchF is but other members of the Obg family are known to be associated with ribosomal Subunits. unknown, Immunoblots of the polysome fraction from sucrose gradients showed that TcYchF was associated with ribosomal subunits and polysomes. Immunoprecipitation assays showed that TcYchF was also associated with the proteasome of T. cruzi. Furthermore, inactivation of the T. brucei homolog of TcYchF by RNA interference inhibited the growth of procyclic forms of the parasite. These data suggest that this protein plays an important role in the translation machinery of trypanosomes. (C) 2008 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.