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一个类似增强子的区域调控人类疟原虫恶性疟原虫中hrp3启动子的阶段特异性基因表达。

An enhancer-like region regulates hrp3 promoter stage-specific gene expression in the human malaria parasite Plasmodium falciparum.

作者信息

López-Estraño Carlos, Gopalakrishnan Anusha M, Semblat Jean-Philippe, Fergus M Ross, Mazier Dominique, Haldar Kasturi

机构信息

Department of Biology, Life Sciences Bldg. Room 409B, The University of Memphis, 3774 Walker Ave. Memphis, Tennessee 38152, USA.

出版信息

Biochim Biophys Acta. 2007 Jul-Aug;1769(7-8):506-13. doi: 10.1016/j.bbaexp.2007.04.009. Epub 2007 May 8.

Abstract

The asexual blood stage of Plasmodium falciparum is comprised of morphologically distinct ring, trophozoite and schizont stages. Each of these developmental stages possesses a distinct pattern of gene expression. Regulation of P. falciparum gene expression is thought to occur, at least in part, at the promoter level. Previously, we have found that although the hrp3 mRNA is only seen in ring-stage parasites, deletion of a specific sequence in the 5' end of the promoter region decreased ring-stage expression of hrp3 and enabled detection of its transcripts in trophozoite-stage parasites. In order to investigate this stage specific regulation of gene expression, we employed a series of nested deletions of the 1.7-kb hrp3 promoter. Firefly luciferase gene was used as a reporter to evaluate the role of promoter sequences in gene regulation. Using this approach, we identified a ring-stage specific regulatory region on the hrp3 promoter located between -1.7 kb and -1.1 kb from the ATG initiation codon. Small 100-150 bp truncations on this enhancer-like region failed to uncover discrete regulatory sequences, suggesting the multipartite nature of this element. The data presented in this study demonstrate that stage specific promoter activity of the hrp3 gene in P. falciparum blood stage parasites is supported, at least in-part, by a small promoter region that can function in the absence of a larger chromosomal context.

摘要

恶性疟原虫的无性血液阶段由形态上不同的环状体、滋养体和裂殖体阶段组成。这些发育阶段中的每一个都具有独特的基因表达模式。恶性疟原虫基因表达的调控至少部分发生在启动子水平。此前,我们发现尽管hrp3 mRNA仅在环状体阶段的寄生虫中可见,但启动子区域5'端特定序列的缺失降低了hrp3在环状体阶段的表达,并使得能够在滋养体阶段的寄生虫中检测到其转录本。为了研究这种基因表达的阶段特异性调控,我们对1.7 kb的hrp3启动子进行了一系列嵌套缺失。萤火虫荧光素酶基因用作报告基因,以评估启动子序列在基因调控中的作用。使用这种方法,我们在hrp3启动子上鉴定出一个环状体阶段特异性调控区域,位于距ATG起始密码子-1.7 kb至-1.1 kb之间。对这个类似增强子的区域进行100 - 150 bp的小片段截短未能发现离散的调控序列,这表明该元件具有多部分性质。本研究中呈现的数据表明,恶性疟原虫血液阶段寄生虫中hrp3基因的阶段特异性启动子活性至少部分由一个小的启动子区域支持,该区域在没有更大的染色体背景的情况下也能发挥作用。

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