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在柔嫩艾美耳球虫中,从头核糖体生物合成受转录调控,这取决于其生命周期阶段。

De novo ribosome biosynthesis is transcriptionally regulated in Eimeria tenella, dependent on its life cycle stage.

作者信息

Schaap Dick, Arts Gerben, van Poppel Nicole F J, Vermeulen Arno N

机构信息

Department of Parasitology, Intervet International BV, P.O. Box 31, 5830AA Boxmeer, The Netherlands.

出版信息

Mol Biochem Parasitol. 2005 Feb;139(2):239-48. doi: 10.1016/j.molbiopara.2004.11.011.

Abstract

Protozoan parasites go through various developmental stages during their parasitic life, which requires the expression of different genes. To identify stage specific gene products in Eimeria tenella, a differential screening was performed comparing the intracellular schizont stage with the extracellular oocyst stage. De novo transcripts of 18S-5.8S-26S rRNA transcription units and of two ribosomal proteins (RPL5 and RPL23) were specifically identified in schizonts and were undetectable in oocysts. The stage specific transcription of pre-rRNAs (prior to processing) was confirmed with Northern blot analysis. Since the E. tenella genome contains a repeated gene cluster with an estimated 140 large rRNA transcription units, they all might be similarly regulated. Specific expression of RPL5 and RPL23 in E. tenella schizonts was also confirmed by Northern blotting. Furthermore, an analysis of the E. tenella EST database with 26,705 ESTs showed that 9.5% of all merozoite ESTs and only 0.2% of the sporozoite ESTs encoded ribosomal proteins (RPs). These ESTs encoded 69 different RPs, suggesting that most and possibly all RPs are differentially transcribed in E. tenella. Analysis of EST data from other Coccidia, such as Toxoplasma gondii, indicated a similar stage dependent transcription of RP genes. We conclude that ribosome biosynthesis is transcriptionally regulated in E. tenella and other Coccidia, such that rapidly growing parasite stages utilize much of their resources to de novo biosynthesis of ribosomes, and that "dormant" oocyst stages do not synthesize new ribosomes. The 50- to 100-fold reduction in transcription of RPs together with the reduced rRNA transcription prevents that unnecessary new ribosomes are synthesized in oocysts.

摘要

原生动物寄生虫在其寄生生活中会经历不同的发育阶段,这需要表达不同的基因。为了鉴定柔嫩艾美耳球虫中的阶段特异性基因产物,进行了一项差异筛选,比较了细胞内裂殖体阶段和细胞外卵囊阶段。在裂殖体中特异性鉴定出了18S - 5.8S - 26S rRNA转录单元以及两种核糖体蛋白(RPL5和RPL23)的从头转录本,而在卵囊中未检测到。通过Northern印迹分析证实了前体rRNA(加工前)的阶段特异性转录。由于柔嫩艾美耳球虫基因组包含一个估计有140个大rRNA转录单元的重复基因簇,它们可能都受到类似的调控。通过Northern印迹法也证实了RPL5和RPL23在柔嫩艾美耳球虫裂殖体中的特异性表达。此外,对含有26,705个EST的柔嫩艾美耳球虫EST数据库进行分析表明,所有裂殖子EST中有9.5%编码核糖体蛋白(RPs),而子孢子EST中只有0.2%编码核糖体蛋白。这些EST编码了69种不同的RPs,表明大多数甚至可能所有的RPs在柔嫩艾美耳球虫中都是差异转录的。对来自其他球虫(如刚地弓形虫)的EST数据进行分析表明,RP基因存在类似的阶段依赖性转录。我们得出结论,核糖体生物合成在柔嫩艾美耳球虫和其他球虫中受到转录调控,使得快速生长的寄生虫阶段将其大部分资源用于核糖体的从头生物合成,而“休眠”的卵囊阶段不合成新的核糖体。核糖体蛋白转录减少50至100倍以及rRNA转录减少,可防止卵囊中合成不必要的新核糖体。

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