In Vivo and In Vitro Characterization of a Plasmodium Liver Stage-Specific Promoter

Niz, Mariana De; Helm, Susanne; Horstmann, Sebastian; Annoura, Takeshi; Portillo, Hernando A. del; Khan, Shahid M.; Heussler, Volker

doi:10.1371/journal.pone.0123473

Cited by 17 publications

(15 citation statements)

References 51 publications

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“…Why expression of sbp1 and mahrp1a and mahrp1b appears to be lower in EF stage merozoites than in EEF stage merozoites is unknown. One reason might be, that in course of EF stage infection, the spleen is heavily remodeled (63–65) and allows for passage of infected RBC, making an efficient sequestration and thus a pronounced expression of sequestration ligands on the surface of the infected RBC less necessary (50). The parasite might thus be less dependent on intense trafficking of sequestration ligands.…”

Section: Resultsmentioning

confidence: 99%

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Transcriptome analysis ofPlasmodium bergheiduring exo-erythrocytic development

Caldelari

Dogga

Schmid³

et al. 2019

Preprint

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SummaryThe complex life cycle of malaria parasites requires well-orchestrated stage specific gene expression. In the vertebrate host the parasites grow and multiply by schizogony in two different environments: within erythrocytes and within hepatocytes. Whereas erythrocytic parasites are rather well-studied in this respect, relatively little is known about the exo-erythrocytic stages. In an attempt to fill this gap, we performed genome wide RNA-seq analyses of various exo-erythrocytic stages of Plasmodium berghei including sporozoites, samples from a time-course of liver stage development and detached cells, which contain infectious merozoites and represent the final step in exo-erythrocytic development. The analysis represents the completion of the transcriptome of the entire life cycle of P. berghei parasites with temporal detailed analysis of the liver stage allowing segmentation of the transcriptome across the progression of the life cycle. We have used these RNA-seq data from different developmental stages to cluster genes with similar expression profiles, in order to infer their functions. A comparison with published data of other parasite stages confirmed stage-specific gene expression and revealed numerous genes that are expressed differentially in blood and exo-erythrocytic stages. One of the most exo-erythrocytic stage-specific genes was PBANKA_1003900, which has previously been annotated as a “gametocyte specific protein”. The promoter of this gene drove high GFP expression in exo-erythrocytic stages, confirming its expression profile seen by RNA-seq. The comparative analysis of the genome wide mRNA expression profiles of erythrocytic and different exo-erythrocytic stages improves our understanding of gene regulation of Plasmodium parasites and can be used to model exo-erythrocytic stage metabolic networks and identify differences in metabolic processes during schizogony in erythrocytes and hepatocytes.

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Section: Resultsmentioning

confidence: 99%

“…6 ). The genes lisp1 (PBANKA_1024600) and lisp2 (PBANKA_1003000) have been previously reported to be expressed exclusively during EEF stage development (65–67). This is clearly reflected by our RNA-seq analysis where substantial lisp1 and lisp2 mRNA levels were only found in EEF stages, from 24hpi to DC.…”

Section: Resultsmentioning

confidence: 99%

Transcriptome analysis ofPlasmodium bergheiduring exo-erythrocytic development

Caldelari

Dogga

Schmid³

et al. 2019

Preprint

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“…In contrast, the uis4 (upregulated in infectious sporozoites) gene is specifically upregulated in infectious sporozoites, those resident in the salivary glands of the mosquitoes and named accordingly [ 32 – 34 ]. Lastly, the lisp2 gene, encoding the liver stage-specific protein 2, is expressed exclusively during late liver stage development prior to formation of infectious merozoites, the parasite forms infecting red blood cells [ 35 ].…”

Section: Resultsmentioning

confidence: 99%

Zinc finger nuclease-based double-strand breaks attenuate malaria parasites and reveal rare microhomology-mediated end joining

et al. 2015

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BackgroundGenome editing of malaria parasites is key to the generation of live attenuated parasites used in experimental vaccination approaches. DNA repair in Plasmodium generally occurs only through homologous recombination. This has been used to generate transgenic parasites that lack one to three genes, leading to developmental arrest in the liver and allowing the host to launch a protective immune response. While effective in principle, this approach is not safe for use in humans as single surviving parasites can still cause disease. Here we use zinc-finger nucleases to generate attenuated parasite lines lacking an entire chromosome arm, by a timed induction of a double-strand break. Rare surviving parasites also allow the investigation of unconventional DNA repair mechanisms in a rodent malaria parasite.ResultsA single, zinc-finger nuclease-induced DNA double-strand break results in the generation of attenuated parasite lines that show varying degrees of developmental arrest, protection efficacy in an immunisation regime and safety, depending on the timing of zinc-finger nuclease expression within the life cycle. We also identify DNA repair by microhomology-mediated end joining with as little as four base pairs, resulting in surviving parasites and thus breakthrough infections.ConclusionsMalaria parasites can repair DNA double-strand breaks with surprisingly small mini-homology domains located across the break point. Timely expression of zinc-finger nucleases could be used to generate a new generation of attenuated parasite lines lacking hundreds of genes.Electronic supplementary materialThe online version of this article (doi:10.1186/s13059-015-0811-1) contains supplementary material, which is available to authorized users.

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“…These studies also determined that LISP2, named after its expression pattern comparable with LISP1 (Tarun et al, 2008), is exclusively expressed in the mid-to-late liver stages stages. Another group has independently confirmed these results by characterizing stage-specific expression using the promoter region of P. berghei LISP2, in conjunction with a dual luminescence system (Helm et al, 2010; De Niz et al, 2015). Immunofluorescence assays of liver stages showed that LISP2 is transported to the vacuolar space and then, after processing, exported into the hepatocyte cytoplasm (Orito et al, 2013; Itani et al, 2014).…”

Section: 6-cys Proteins In the Pre-erythrocytic Stagesmentioning

confidence: 88%

The s48/45 six-cysteine proteins: mediators of interaction throughout the Plasmodium life cycle

Arredondo

Kappe

2017

International Journal for Parasitology

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During their life cycle Plasmodium parasites rely upon an arsenal of proteins that establish key interactions with the host or vector, and between the parasite sexual stages, with the purpose of ensuring infection, reproduction and proliferation. Among these is a group of secreted or membrane-anchored proteins known as the six-cysteine (6-cys) family. This is a small but important family with only 14 members thus far identified, each stage-specifically expressed during the parasite life cycle. 6-cys proteins often localize at the parasite surface or interface with the host and are conserved in different Plasmodium species. The unifying feature of the family is the s48/45 domain, presumably involved in adhesion and structurally related to Ephrins, the ligands of Eph receptors. The most prominent s48/45 members are currently under functional investigation and are being pursued as vaccine candidates. In this review, we examine what is known about the 6-cys family, their structure and function, and discuss future research directions.

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In Vivo and In Vitro Characterization of a Plasmodium Liver Stage-Specific Promoter

Cited by 17 publications

References 51 publications

Transcriptome analysis ofPlasmodium bergheiduring exo-erythrocytic development

Transcriptome analysis ofPlasmodium bergheiduring exo-erythrocytic development

Zinc finger nuclease-based double-strand breaks attenuate malaria parasites and reveal rare microhomology-mediated end joining

The s48/45 six-cysteine proteins: mediators of interaction throughout the Plasmodium life cycle

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