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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Plasmodium falciparum XPD translocates in 5' to 3' direction, is expressed throughout the blood stages, and interacts with p44
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Plasmodium falciparum XPD translocates in 5' to 3' direction, is expressed throughout the blood stages, and interacts with p44

机译：恶性疟原虫XPD沿5'到3'方向移位，在整个血液阶段表达，并与p44相互作用

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摘要

XPD helicase, a TFIIH subunit, is essential for several processes including transcription, NER, cell cycle regulation, and apoptosis in eukaryotes. Another component of TFIIH, namely p44, is among the well-known interacting partners of XPD and is vital in regulating the helicase activities of latter. However, none of the above mentioned proteins have been functionally characterized in Plasmodium falciparum. Consequently, in this study, we performed detailed studies on XPD and its interacting partner, p44, from P. falciparum 3D7 strain. Accordingly, we expressed and purified recombinant PfXPD and its fragments and Pfp44 proteins and characterized the enzymatic activities of PfXPD and its fragments. The in vivo stage-specific expression and subcellular localizations of PfXPD and Pfp44 proteins were studied using the specific antibodies in the intraerythrocytic developmental stages of P. falciparum 3D7 strain. Our results suggest that PfXPD displays the characteristic ssDNA-dependent ATPase and 5'aEuro"3' DNA helicase activities. We also report the existence of two high molecular weight forms of p44 in P. falciparum 3D7 strain. Both PfXPD and Pfp44 colocalize in the nucleus and interact with each other, which suggest that they are most likely components of the same complex apparently, TFIIH. Furthermore, during trophozoite and schizont stages, both proteins exhibit a distinct cytoplasmic distribution pattern which implies that PfXPD and Pfp44 might also be involved in other functions. These studies will aid in understanding the basic biology of malaria parasite.
机译：XPD解旋酶（一种TFIIH亚基）对于包括转录，NER，细胞周期调控和真核细胞凋亡在内的多个过程至关重要。 TFIIH的另一种成分，即p44，是XPD众所周知的相互作用伴侣，在调节XPD的解旋酶活性中至关重要。但是，上述蛋白均未在恶性疟原虫中进行功能鉴定。因此，在这项研究中，我们对恶性疟原虫3D7菌株的XPD及其相互作用伴侣p44进行了详细研究。因此，我们表达和纯化了重组PfXPD及其片段和Pfp44蛋白，并表征了PfXPD及其片段的酶活性。在恶性疟原虫3D7菌株的红细胞内发育阶段，使用特异性抗体研究了PfXPD和Pfp44蛋白的体内阶段特异性表达和亚细胞定位。我们的结果表明，PfXPD表现出特征性的ssDNA依赖性ATPase和5'aEuro“ 3'DNA解旋酶活性。我们还报道了恶性疟原虫3D7菌株中存在两种高分子量形式的p44，PfXPD和Pfp44共同定位在恶性疟原虫中。核彼此相互作用，这表明它们很可能是同一复合物TFIIH的最可能组成部分;此外，在滋养体和裂殖体阶段，两种蛋白均表现出独特的胞质分布模式，这表明PfXPD和Pfp44也可能参与其中。这些研究将有助于理解疟疾寄生虫的基本生物学。

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