Initial characterization of the Pf-Int recombinase from the malaria parasite Plasmodium falciparum.
Initial characterization of the Pf-Int recombinase from the malaria parasite Plasmodium falciparum.
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DOI：
10.1371/journal.pone.0046507
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Gopaul DN
中科院分区：
文献类型：
--
作者：
Ghorbal M;Scheidig-Benatar C;Bouizem S;Thomas C;Paisley G;Faltermeier C;Liu M;Scherf A;Lopez-Rubio JJ;Gopaul DN
遗传变异是许多生物为应对环境变化而进行进化和适应的重要手段。某些DNA改变可以由位点特异性重组酶（SSRs）进行，它分为两个家族：丝氨酸重组酶和酪氨酸重组酶。在真核生物中很少发现SSRs。在恶性疟原虫基因组中发现了一个与酪氨酸位点特异性重组酶同源的基因。该序列在疟原虫的其他5个成员中高度保守。预测的开放阅读框编码一个约57 kDa的蛋白，该蛋白含有一个c端结构域，包括假定的酪氨酸重组酶保守活性位点残基R-H-R-(H/W)- y。n端具有典型的α -螺旋束和潜在的类似于λ-Int的混合α - β结构域。Pf-Int mRNA在恶性疟原虫红细胞生命阶段表达差异，在分裂期达到峰值。重组Pf-Int和亲和层析的DNA从基因组或合成来源，以确定测序或微阵列杂交后潜在的DNA靶点。有趣的是，捕获的序列还包括高度可变的亚端粒基因，如var、rif和stevor序列。用DNA进行电泳迁移转移试验以验证Pf-Int/DNA结合。最后，为了研究Pf-Int的生物学作用，构建了Pf-Int敲除寄生虫。我们的数据首次鉴定出具有重组酶结构和功能特征的疟原虫基因。Pf-Int可以以序列特异性或非特异性的方式结合和改变DNA，并可能有助于程序化或随机的DNA重排。Pf-Int是第一个在该病原体基因组可塑性中具有潜在作用的分子参与者。最后，Pf-Int敲除寄生虫是有活力的，对血液阶段的发育没有可检测到的影响，这与这种功能是兼容的。
Genetic variation is an essential means of evolution and adaptation in many organisms in response to environmental change. Certain DNA alterations can be carried out by site-specific recombinases (SSRs) that fall into two families: the serine and the tyrosine recombinases. SSRs are seldom found in eukaryotes. A gene homologous to a tyrosine site-specific recombinase has been identified in the genome of Plasmodium falciparum. The sequence is highly conserved among five other members of Plasmodia. The predicted open reading frame encodes for a ∼57 kDa protein containing a C-terminal domain including the putative tyrosine recombinase conserved active site residues R-H-R-(H/W)-Y. The N-terminus has the typical alpha-helical bundle and potentially a mixed alpha-beta domain resembling that of λ-Int. Pf-Int mRNA is expressed differentially during the P. falciparum erythrocytic life stages, peaking in the schizont stage. Recombinant Pf-Int and affinity chromatography of DNA from genomic or synthetic origin were used to identify potential DNA targets after sequencing or micro-array hybridization. Interestingly, the sequences captured also included highly variable subtelomeric genes such as var, rif, and stevor sequences. Electrophoretic mobility shift assays with DNA were carried out to verify Pf-Int/DNA binding. Finally, Pf-Int knock-out parasites were created in order to investigate the biological role of Pf-Int. Our data identify for the first time a malaria parasite gene with structural and functional features of recombinases. Pf-Int may bind to and alter DNA, either in a sequence specific or in a non-specific fashion, and may contribute to programmed or random DNA rearrangements. Pf-Int is the first molecular player identified with a potential role in genome plasticity in this pathogen. Finally, Pf-Int knock-out parasite is viable showing no detectable impact on blood stage development, which is compatible with such function.
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DOI：
10.1038/nrmicro2145
发表时间：
2009-07
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
通讯作者：
--
影响因子：
11.4
作者：
Bouvier, M;Demarre, G;Mazel, D
通讯作者：
Mazel, D
影响因子：
64.8
作者：
Freitas-Junior, LH;Bottius, E;Scherf, A
通讯作者：
Scherf, A
影响因子：
64.8
作者：
Guo, F;Gopaul, DN;VanDuyne, GD
通讯作者：
VanDuyne, GD
影响因子：
3.4
作者：
Brown, Patrick H.;Schuck, Peter
通讯作者：
Schuck, Peter