Identification and characterization of a liver stage-specific promoter region of the malaria parasite plasmodium

Susanne Helm, Christine Lehmann, Andreas Nagel, Rebecca R. Stanway, Sebastian Horstmann, Manuel Llinas, Volker T. Heussler

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

During the blood meal of a Plasmodium-infected mosquito, 10 to 100 parasites are inoculated into the skin and a proportion of these migrate via the bloodstream to the liver where they infect hepatocytes. The Plasmodium liver stage, despite its clinical silence, represents a highly promising target for antimalarial drug and vaccine approaches. Successfully invaded parasites undergo a massive proliferation in hepatocytes, producing thousands of merozoites that are transported into a blood vessel to infect red blood cells. To successfully develop from the liver stage into infective merozoites, a tight regulation of gene expression is needed. Although this is a very interesting aspect in the biology of Plasmodium, little is known about gene regulation in Plasmodium parasites in general and in the liver stage in particular. We have functionally analyzed a novel promoter region of the rodent parasite Plasmodium berghei that is exclusively active during the liver stage of the parasite. To prove stage-specific activity of the promoter, GFP and luciferase reporter assays have been successfully established, allowing both qualitative and accurate quantitative analysis. To further characterize the promoter region, the transcription start site was mapped by rapid amplification of cDNA ends (5′-RACE). Using promoter truncation experiments and site-directed mutagenesis within potential transcription factor binding sites, we suggest that the minimal promoter contains more than one binding site for the recently identified parasite-specific ApiAP2 transcription factors. The identification of a liver stage-specific promoter in P. berghei confirms that the parasite is able to tightly regulate gene expression during its life cycle. The identified promoter region might now be used to study the biology of the Plasmodium liver stage, which has thus far proven problematic on a molecular level. Stage-specific expression of dominant-negative mutant proteins and over expression of proteins normally active in other life cycle stages will help to understand the function of the proteins investigated.

Original languageEnglish (US)
Article numbere13653
JournalPloS one
Volume5
Issue number10
DOIs
StatePublished - Nov 17 2010

Fingerprint

Plasmodium malariae
Plasmodium
Genetic Promoter Regions
Liver
malaria
Parasites
promoter regions
parasites
liver
Gene expression
Plasmodium berghei
Merozoites
merozoites
Life Cycle Stages
hepatocytes
Life cycle
binding sites
Hepatocytes
Blood
Transcription Factors

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Helm, Susanne ; Lehmann, Christine ; Nagel, Andreas ; Stanway, Rebecca R. ; Horstmann, Sebastian ; Llinas, Manuel ; Heussler, Volker T. / Identification and characterization of a liver stage-specific promoter region of the malaria parasite plasmodium. In: PloS one. 2010 ; Vol. 5, No. 10.
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abstract = "During the blood meal of a Plasmodium-infected mosquito, 10 to 100 parasites are inoculated into the skin and a proportion of these migrate via the bloodstream to the liver where they infect hepatocytes. The Plasmodium liver stage, despite its clinical silence, represents a highly promising target for antimalarial drug and vaccine approaches. Successfully invaded parasites undergo a massive proliferation in hepatocytes, producing thousands of merozoites that are transported into a blood vessel to infect red blood cells. To successfully develop from the liver stage into infective merozoites, a tight regulation of gene expression is needed. Although this is a very interesting aspect in the biology of Plasmodium, little is known about gene regulation in Plasmodium parasites in general and in the liver stage in particular. We have functionally analyzed a novel promoter region of the rodent parasite Plasmodium berghei that is exclusively active during the liver stage of the parasite. To prove stage-specific activity of the promoter, GFP and luciferase reporter assays have been successfully established, allowing both qualitative and accurate quantitative analysis. To further characterize the promoter region, the transcription start site was mapped by rapid amplification of cDNA ends (5′-RACE). Using promoter truncation experiments and site-directed mutagenesis within potential transcription factor binding sites, we suggest that the minimal promoter contains more than one binding site for the recently identified parasite-specific ApiAP2 transcription factors. The identification of a liver stage-specific promoter in P. berghei confirms that the parasite is able to tightly regulate gene expression during its life cycle. The identified promoter region might now be used to study the biology of the Plasmodium liver stage, which has thus far proven problematic on a molecular level. Stage-specific expression of dominant-negative mutant proteins and over expression of proteins normally active in other life cycle stages will help to understand the function of the proteins investigated.",
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Identification and characterization of a liver stage-specific promoter region of the malaria parasite plasmodium. / Helm, Susanne; Lehmann, Christine; Nagel, Andreas; Stanway, Rebecca R.; Horstmann, Sebastian; Llinas, Manuel; Heussler, Volker T.

In: PloS one, Vol. 5, No. 10, e13653, 17.11.2010.

Research output: Contribution to journalArticle

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