De Silva Erandi K, Gehrke Andrew R, Olszewski Kellen, León Ilsa, Chahal Jasdave S, Bulyk Martha L, Llinás Manuel
Department of Molecular Biology and Lewis-Sigler Institute for Integrative Genomics, Princeton University, 246 Carl Icahn Laboratory, Princeton, NJ 08544, USA.
Proc Natl Acad Sci U S A. 2008 Jun 17;105(24):8393-8. doi: 10.1073/pnas.0801993105. Epub 2008 Jun 9.
Malaria remains one of the most prevalent infectious diseases worldwide, affecting more than half a billion people annually. Despite many years of research, the mechanisms underlying transcriptional regulation in the malaria-causing Plasmodium spp., and in Apicomplexan parasites generally, remain poorly understood. In Plasmodium, few regulatory elements sufficient to drive gene expression have been characterized, and their cognate DNA-binding proteins remain unknown. This study characterizes the DNA-binding specificities of two members of the recently identified Apicomplexan AP2 (ApiAP2) family of putative transcriptional regulators from Plasmodium falciparum. The ApiAP2 proteins contain AP2 domains homologous to the well characterized plant AP2 family of transcriptional regulators, which play key roles in development and environmental stress response pathways. We assayed ApiAP2 protein-DNA interactions using protein-binding microarrays and combined these results with computational predictions of coexpressed target genes to couple these putative trans factors to corresponding cis-regulatory motifs in Plasmodium. Furthermore, we show that protein-DNA sequence specificity is conserved in orthologous proteins between phylogenetically distant Apicomplexan species. The identification of the DNA-binding specificities for ApiAP2 proteins lays the foundation for the exploration of their role as transcriptional regulators during all stages of parasite development. Because of their origin in the plant lineage, ApiAP2 proteins have no homologues in the human host and may prove to be ideal antimalarial targets.
疟疾仍然是全球最普遍的传染病之一,每年影响着超过5亿人。尽管经过多年研究,但导致疟疾的疟原虫属以及一般顶复门寄生虫中转录调控的潜在机制仍知之甚少。在疟原虫中,很少有足以驱动基因表达的调控元件得到表征,其同源DNA结合蛋白也仍然未知。本研究表征了来自恶性疟原虫的最近鉴定出的顶复门AP2(ApiAP2)家族两个假定转录调节因子成员的DNA结合特异性。ApiAP2蛋白含有与特征明确的植物转录调节因子AP2家族同源的AP2结构域,这些结构域在发育和环境应激反应途径中起关键作用。我们使用蛋白质结合微阵列分析了ApiAP2蛋白与DNA的相互作用,并将这些结果与共表达靶基因的计算预测相结合,以将这些假定的转录因子与疟原虫中的相应顺式调控基序联系起来。此外,我们表明,在系统发育上距离较远的顶复门物种之间,直系同源蛋白中的蛋白质与DNA序列特异性是保守的。鉴定ApiAP2蛋白的DNA结合特异性为探索其在寄生虫发育各个阶段作为转录调节因子的作用奠定了基础。由于它们起源于植物谱系,ApiAP2蛋白在人类宿主中没有同源物,可能被证明是理想的抗疟靶点。