Transcriptome analysis reveals unique metabolic features in the Cryptosporidium parvum Oocysts associated with environmental survival and stresses.
Transcriptome analysis reveals unique metabolic features in the Cryptosporidium parvum Oocysts associated with environmental survival and stresses.
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DOI：
10.1186/1471-2164-13-647
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发表时间：
2012-11-21
期刊：
影响因子：
4.4
通讯作者：
Zhu G
中科院分区：
文献类型：
--
作者：
Zhang H;Guo F;Zhou H;Zhu G
微小隐孢子虫是一种全球性分布的人畜共患寄生虫，也是免疫功能低下患者的重要机会致病菌。对寄生虫的代谢动力学知之甚少，并且由于缺乏分子和遗传工具，研究受到阻碍。在这里，我们报告的发展，第一安捷伦芯片的C。parvum（CpArray 15 K），其覆盖寄生虫基因组中所有预测的ORF。使用CpArray 15 K结合实时qRT-PCR的全局转录组分析揭示了卵囊中的许多独特代谢特征，卵囊是寄生虫的感染和环境阶段。基于与核糖体生物合成、转录和翻译相关的基因的高转录水平，发现卵囊期寄生虫在蛋白质合成中高度活跃。蛋白酶体和泛素相关的组件也是高度活跃的，这意味着卵囊可能采用蛋白质降解途径回收氨基酸，以克服不能从头合成氨基酸。乳酸脱氢酶（LDH）基因在卵囊中表达量最高，是卵囊能量代谢的特征。我们还研究了寄生虫对紫外线照射的反应，并观察到基因表达的复杂和动态调节。值得注意的变化包括参与DNA修复和细胞内运输的基因的转录水平增加。在应激相关基因中，TCP-1家族成员和硫氧还蛋白相关基因在卵囊紫外线损伤后的恢复过程中起着重要作用。我们的观察结果还表明，紫外线照射卵囊的结果在细胞骨架重排和细胞内膜贩运的活动增加。CpArray 15 K是第一个针对C. parvum，为隐孢子虫研究界提供了一个研究寄生虫转录组和功能基因组学所需的工具。CpArray 15 K已成功地用于分析寄生虫卵囊中的基因表达以及它们对紫外线照射的反应。这些观察结果揭示了寄生虫卵囊如何适应和应对不利的外部环境和相关的压力，如紫外线照射。
Cryptosporidium parvum is a globally distributed zoonotic parasite and an important opportunistic pathogen in immunocompromised patients. Little is known on the metabolic dynamics of the parasite, and study is hampered by the lack of molecular and genetic tools. Here we report the development of the first Agilent microarray for C. parvum (CpArray15K) that covers all predicted ORFs in the parasite genome. Global transcriptome analysis using CpArray15K coupled with real-time qRT-PCR uncovered a number of unique metabolic features in oocysts, the infectious and environmental stage of the parasite. Oocyst stage parasites were found to be highly active in protein synthesis, based on the high transcript levels of genes associated with ribosome biogenesis, transcription and translation. The proteasome and ubiquitin associated components were also highly active, implying that oocysts might employ protein degradation pathways to recycle amino acids in order to overcome the inability to synthesize amino acids de novo. Energy metabolism in oocysts was featured by the highest level of expression of lactate dehydrogenase (LDH) gene. We also studied parasite responses to UV-irradiation, and observed complex and dynamic regulations of gene expression. Notable changes included increased transcript levels of genes involved in DNA repair and intracellular trafficking. Among the stress-related genes, TCP-1 family members and some thioredoxin-associated genes appear to play more important roles in the recovery of UV-induced damages in the oocysts. Our observations also suggest that UV irradiation of oocysts results in increased activities in cytoskeletal rearrangement and intracellular membrane trafficking. CpArray15K is the first microarray chip developed for C. parvum, which provides the Cryptosporidium research community a needed tool to study the parasite transcriptome and functional genomics. CpArray15K has been successfully used in profiling the gene expressions in the parasite oocysts as well as their responses to UV-irradiation. These observations shed light on how the parasite oocysts might adapt and respond to the hostile external environment and associated stress such as UV irradiation.
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