Identification of a novel and unique transcription factor in the intraerythrocytic stage of Plasmodium falciparum.
Identification of a novel and unique transcription factor in the intraerythrocytic stage of Plasmodium falciparum.
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DOI：
10.1371/journal.pone.0074701
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Kano S
中科院分区：
文献类型：
--
作者：
Komaki-Yasuda K;Okuwaki M;Nagata K;Kawazu S;Kano S
恶性疟原虫阶段特异性基因调控的机制在很大程度上是不清楚的，只有少数特定的调节转录因子（AP 2家族）已被确定。特别是，在红细胞内阶段发挥作用的转录因子仍有待阐明。作为恶性疟原虫红细胞内阶段特异性转录的模型案例，我们分析了滋养体/滋养体特异性基因pf 1-cys-prx的转录调控，并提出一些核因子特异性结合pf 1-cys-prx的顺式元件并增强转录。本研究采用5步层析法从寄生虫核提取物中分离纯化了核因子，并鉴定出一种名为PREBP的因子。PREBP不包括在AP 2家族中，并且是具有四个K-同源（KH）结构域的新蛋白。已知KH结构域存在于RNA结合蛋白或单链DNA结合蛋白中。PREBP在疟原虫属中高度保守，在顶复门中部分保守。为了评估PREBP过表达的影响，我们使用了瞬时过表达和荧光素酶测定相结合的方法。在pf 1-cys-prx顺式元件的控制下，PREBP的过表达显著增强了荧光素酶的表达。这些结果提供了第一个证据，一种新的转录因子，激活疟原虫红细胞内阶段的基因表达。这些发现增强了我们对疟原虫和其他真核生物中特定转录机制进化的理解。
The mechanisms of stage-specific gene regulation in the malaria parasite Plasmodium falciparum are largely unclear, with only a small number of specific regulatory transcription factors (AP2 family) having been identified. In particular, the transcription factors that function in the intraerythrocytic stage remain to be elucidated. Previously, as a model case for stage-specific transcription in the P. falciparum intraerythrocytic stage, we analyzed the transcriptional regulation of pf1-cys-prx, a trophozoite/schizont-specific gene, and suggested that some nuclear factors bind specifically to the cis-element of pf1-cys-prx and enhance transcription. In the present study, we purified nuclear factors from parasite nuclear extract by 5 steps of chromatography, and identified a factor termed PREBP. PREBP is not included in the AP2 family, and is a novel protein with four K-homology (KH) domains. The KH domain is known to be found in RNA-binding or single-stranded DNA-binding proteins. PREBP is well conserved in Plasmodium species and partially conserved in phylum Apicomplexa. To evaluate the effects of PREBP overexpression, we used a transient overexpression and luciferase assay combined approach. Overexpression of PREBP markedly enhanced luciferase expression under the control of the pf1-cys-prx cis-element. These results provide the first evidence of a novel transcription factor that activates the gene expression in the malaria parasite intraerythrocytic stage. These findings enhance our understanding of the evolution of specific transcription machinery in Plasmodium and other eukaryotes.
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