ANK1 and DnaK-TPR, Two Tetratricopeptide Repeat-Containing Proteins Primarily Expressed in Toxoplasma Bradyzoites, Do Not Contribute to Bradyzoite Differentiation.
ANK1 and DnaK-TPR, Two Tetratricopeptide Repeat-Containing Proteins Primarily Expressed in Toxoplasma Bradyzoites, Do Not Contribute to Bradyzoite Differentiation.
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ANK1 和 DnaK-TPR，两种主要在弓形虫缓殖子中表达的含有四肽重复序列的蛋白质，对缓殖子分化没有贡献
DOI：
10.3389/fmicb.2017.02210
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发表时间：
2017
影响因子：
5.2
通讯作者：
Shen B
中科院分区：
文献类型：
--
作者：
Yang J;Zhang L;Diao H;Xia N;Zhou Y;Zhao J;Shen B
弓形虫是一种重要的人畜共患病病原体，感染了世界三分之一的人口和众多的动物。它广泛分布的一个关键因素是能够在快速复制的速殖子和缓慢生长的慢殖子之间相互转换，并在中间宿主中建立终身慢性感染的能力。虽然人们普遍认为阶段转换在寄生虫的发病和传播中起着关键作用，但对其背后的分子机制知之甚少。利用TOXODB现有的基因表达数据和已发表的工作，我们寻找具有新的功能结构域的蛋白质，这些蛋白质在缓体阶段表达上调，希望找到在阶段转换和缓体形成过程中具有关键作用的分子。在这项研究中，我们鉴定了两个这样的蛋白ANK1和DNAK-TPR，这两个蛋白都主要在缓殖子中表达，并且都含有新的基序来调节蛋白质之间的相互作用。通过CRISPR/Cas9定向基因编辑技术，分别敲除了1型TgHB2株和2型ME49株的两个基因。这两个基因的破坏都不会影响速殖子在体外的生长或复制，这与它们在这个阶段的最低表达是一致的。然而，在小鼠感染过程中，缺乏ANK1或DNAK-TPR的突变体显示出轻微的毒力减弱。令人惊讶的是，ANK1和DNAK-TPR的失活似乎都对体外缓殖子分化或体内包囊形成有显著影响。这些结果表明，ANK1和DNAK-TPR可能不直接促进缓殖体分化，但可能影响缓殖体生物学的其他方面。
Toxoplasma gondii is an important zoonotic pathogen infecting one third of the world population and numerous animals. A key factor to its wide distribution is the ability to interconvert between fast replicating tachyzoites and slowly growing bradyzoites, and to establish lifelong chronic infection in intermediate hosts. Although it is well accepted that stage conversion plays key roles in the pathogenesis and transmission of the parasite, little is known about the molecular mechanisms behind it. Using existing gene expression data from TOXODB and published work, we looked for proteins with novel functional domains and whose expression is up-regulated in the bradyzoite stage, hoping to find molecules that have critical roles in regulating stage conversion and bradyzoite formation. In this study we characterized two such proteins ANK1 and DnaK-TPR, both of which are primarily expressed in bradyzoites and contain novel motifs to mediate protein-protein interactions. Through CRISPR/CAS9 directed gene editing technology, both genes were individually knocked out in type 1 strain TgHB2 and type 2 strain ME49. Disruption of neither of these two genes affected the growth or replication of tachyzoites in vitro, consistent with their minimal expression at this stage. However, mutants lacking ANK1 or DnaK-TPR displayed modest virulence attenuation during mice infection. Surprisingly, inactivation of neither ANK1 nor DnaK-TPR seemed to have a significant impact on bradyzoite differentiation in vitro or cyst formation in vivo. These results suggest that ANK1 and DnaK-TPR probably do not directly contribute to bradyzoite differentiation, but likely affect other aspects of bradyzoite biology.
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DOI：
10.3347/kjp.2012.50.3.199
发表时间：
2012-09
期刊：
The Korean journal of parasitology
影响因子：
--
作者：
Selseleh M;Modarressi MH;Mohebali M;Shojaee S;Eshragian MR;Selseleh M;Azizi E;Keshavarz H
通讯作者：
Keshavarz H
影响因子：
3.7
作者：
Abdelbaset AE;Fox BA;Karram MH;Abd Ellah MR;Bzik DJ;Igarashi M
通讯作者：
Igarashi M
影响因子：
4
作者：
Dubey, JP
通讯作者：
Dubey, JP
影响因子：
--
作者：
Buchholz, Kerry R.;Bowyer, Paul W.;Boothroyd, John C.
通讯作者：
Boothroyd, John C.
影响因子：
3.1
作者：
Manger, ID;Hehl, A;Boothroyd, JC
通讯作者：
Boothroyd, JC