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Comparative transcriptional analysis of pre-erythrocytic stages

ABSTRACT: To gain insight into the molecular mechanisms at work during progression through the pre-erythrocytic stages, a comparative microarray based transcriptional study was under taken on radiation attenuated (RAS) and wild type sporozoites (wtSPZ) as well as, and liver stage parasites collected 24 hours (24hrLS) and 48 hours (48hrLS) after wild type sporozoite infection. We were able to identify ~1100 genes significantly differentially expressed during one or more of the pre-erythrocytic stages relative to the mixed blood stages.

ORGANISM(S): Plasmodium yoelii

PROVIDER: GSE18730 | GEO | 2009/11/10

SECONDARY ACCESSION(S): PRJNA121633

REPOSITORIES: GEO

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Comparative transcriptional analysis of pre-erythrocytic stages

Project description:To gain insight into the molecular mechanisms at work during progression through the pre-erythrocytic stages, a comparative microarray based transcriptional study was under taken on radiation attenuated (RAS) and wild type sporozoites (wtSPZ) as well as, and liver stage parasites collected 24 hours (24hrLS) and 48 hours (48hrLS) after wild type sporozoite infection. We were able to identify ~1100 genes significantly differentially expressed during one or more of the pre-erythrocytic stages relative to the mixed blood stages. This study compared the gene expression profiles of radiation attenuated sporozoites (RAS), wild type sporooites (wtSPZ), and infected hepatocytes collected 24hr (24hrLS) and 48hr (48hrLS) after sporozoite infection, using a common reference design. Each sample was hybridized with an equal amount of reference RNA. 48hrLS: 4 biological replicates, with 2 total technical replicates and one dye swap; 24hrLS: 4 biologicla replicates, with 3 total technical replicates; RAS: 2 biological replicates, with 2 technical replicates total, and two tota dye swaps; wtSPZ: 2 biological replicates, with 2 total technical replicates and 2 total dye swaps

2010-06-20 | E-GEOD-18730 | biostudies-arrayexpress

Plasmodium berghei PbeIK2KO sporozoites

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Transcription profiling by array of Plasmodium berghei wild type and puf2 knock out in a time series

Project description:Many eukaryotic developmental and cell fate decisions are effected post-transcriptionally that mechanistically involve RNA binding proteins as regulators of translation of key mRNAs. In the unicellular eukaryote malaria parasite, Plasmodium, one of the most dramatic changes in cell morphology and function occurs during transmission between mosquito and human host. In the mosquito salivary glands, Plasmodium sporozoites are slender, motile and remain infectious for several weeks; only after transmission and liver cell invasion, does the parasite rapidly transform into a round, non-motile exo-erythrocytic form (EEF) that gives rise to thousands of infectious merozoites to be released into the blood stream. Here we demonstrate a Plasmodium homolog of the RNA binding protein, Pumilio, as a key regulator of the sporozoite to EEF transformation. In the absence of Pumilio-2 (Puf2) Plasmodium berghei sporozoites initiate early stage EEF development inside mosquito salivary glands with characteristic morphological changes; puf2- salivary gland sporozoites lose gliding motility, cell traversal ability and are less infective. Global expression profiling confirmed that transgenic parasites exhibit genome-wide transcriptional adaptations typical for Plasmodium intra-hepatic development. The data demonstrate that Puf2 is a key player in regulating developmental control, and imply that transformation of salivary gland-resident sporozoites into early liver stage parasites is regulated by a post-translational mechanism.

2010-11-01 | E-TABM-1067 | biostudies-arrayexpress

Plasmodium berghei PbeIK2KO sporozoites

Project description:Malaria sporozoites, the form transmitted by mosquitoes, are quiescent while in the insect salivary glands. It is only after the sporozoites are deposited in the host’s skin, migrate to the liver and infect hepatocytes that the parasites continue the life cycle. We show that the sporozoite latency is an active process that requires phosphorylation of the eukaryotic initiation factor-2α (eIF2α) by a sporozoite-specific kinase. Inactivation of the kinase gene leads to an overall enhancement of protein synthesis including of silenced liver stage proteins, and inhibits transmission of malaria. Specific inhibition of the eIF2α phosphatase by salubrinal has the opposite effect. Thus, to prevent premature transformation into liver stages, Plasmodium sporozoites exploit the same mechanism that regulates stress responses in mammalian cells.

2009-07-01 | GSE16259 | GEO

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Project description:Plasmodium berghei transcriptome for female gametocytes, male gametocytes, and asexual erythrocytic stages

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Regulation of Puf2 Expression is Critical for Plasmodium Sporozoite Development and Maintenance of Sporozoite Transmissibility

Project description:MalariaM-bM-^@M-^Ys cycle of infection requires parasite transmission between a mosquito vector and a vertebrate host. Plasmodium regulates transmission by translationally repressing specific mRNAs until their products are needed. We demonstrate that the Plasmodium yoelii Pumilio-FBF family member Puf2 allows the sporozoite to retain its infectivity in the mosquito salivary glands while awaiting transmission. Puf2 mediates this critical feature solely through its RNA-Binding Domain (RBD) likely by protecting and silencing specific mRNAs. Puf2 storage granules are distinct from stress granules and P-bodies and dissolve rapidly after infection of hepatocytes, likely releasing the protected and silenced transcripts for M-bM-^@M-^Xjust-in-timeM-bM-^@M-^Y translation by early exoerythrocytic forms (EEFs). Further corroborating this model, pypuf2- sporozoites have no apparent defect in host infection early after invading the salivary glands, but become progressively noninfectious and subsequently prematurely transform into EEFs during prolonged salivary gland residence. In contrast, the premature overexpression of Puf2 in oocysts causes striking deregulation of sporozoite maturation, resulting in fewer oocyst sporozoites that are non-infectious and unable to colonize the salivary glands. Maintenance of Puf2 expression in liver stage parasites produces no phenotype, suggesting that a window of permissive expression exists. Finally, by conducting the first comparative RNAseq analysis of Plasmodium sporozoites, we have uncovered that Puf2 may play a role in both the protection of specific transcripts as well as RNA turnover via the CCR4/Not complex. These findings uncover requirements for maintaining a window of opportunity for the malaria parasite to accommodate the unpredictable moment of transmission from mosquito to vertebrate host. Wild-type (Py17XNL) and pypuf2 -salivary gland sporozoites

2012-10-27 | E-GEOD-41873 | biostudies-arrayexpress

Plasmodium yoelii

Project description:Comparative transcriptional analysis of pre-erythrocytic stages

| PRJNA121633 | ENA

In-depth proteomic analysis of Plasmodium berghei sporozoites using trapped ion mobility spectrometry with parallel accumulation-serial fragmentation

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Transcription profiling of wild type, Pfsir2A knock-out and Pfsir2B knock-out Plasmodium falciparum intra-erythrocytic stages

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Comparative Total RNA-seq between WT and alba4-null Plasmodium yoelii parasites

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