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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Interactions of PfETRAMP14.1 with PfEMP1 and EXP-2 at the parasitophorous vacuolar membrane (PVM) of human malaria parasite Plasmodium falciparum [LOW PARASITEMIA ANALYSIS]

ABSTRACT: In our global differential gene expression analyses,ETRAMP14.1, an ETRAMP family member was found to be highly transcribed in vivo in severe Malaria patients from highly endemic Indian region. This study for the first time reports the interaction of ETRAMP14.1 with PfEMP1, EXP2 and Hsp70-1. Therefore, we propose that ETRAMP14.1 facilitate PfEMP1 to cross the PVM via transcolon machinery component EXP2. Peripheral blood samples containing ring forms of the P.falciparum from mild and severe malaria patients were used for RNA extraction, CDNA synthesis and hybridization on Affymetrix microarrays. In vitro ring stage P.falciparum cultures were used for normalization.

ORGANISM(S): Homo sapiens

SUBMITTER: Sanjeev Kumar

PROVIDER: E-GEOD-72447 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Interactions of PfETRAMP14.1 with PfEMP1 and EXP-2 at the parasitophorous vacuolar membrane (PVM) of human malaria parasite Plasmodium falciparum [HIGH PARASITEMIA ANALYSIS]

Project description:In our global differential gene expression analyses,ETRAMP14.1, an ETRAMP family member was found to be highly transcribed in vivo in severe Malaria patients from highly endemic Indian region. This study for the first time reports the interaction of ETRAMP14.1 with PfEMP1, EXP2 and Hsp70-1. Therefore, we propose that ETRAMP14.1 facilitate PfEMP1 to cross the PVM via transcolon machinery component EXP2. Peripheral blood samples containing ring forms of the P.falciparum from mild and severe malaria patients were used for RNA extraction, CDNA synthesis and hybridization on Affymetrix microarrays. In vitro ring stage P.falciparum cultures were used for normalization.

2015-08-28 | E-GEOD-72446 | biostudies-arrayexpress

Interactions of PfETRAMP14.1 with PfEMP1 and EXP-2 at the parasitophorous vacuolar membrane (PVM) of human malaria parasite Plasmodium falciparum

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2015-08-28 | E-GEOD-72448 | biostudies-arrayexpress

Transcriptional profiling of growth perturbations of the human malaria parasite Plasmodium falciparum

Project description:The microarray experiments were carried out using a long oligonucleotide DNA microarray that represent all 5363 P. falciparum genes with one oligonucleotide per 1.9kb of coding sequence on average (Hu et al. 2007). Total 247 microarray experiments were carried out including 29-drug treatment time courses with 20 compounds and corresponding untreated controls from different drug or inhibitor treatment. Data of each drug/inhibitor experiment were normalized using a linear normalization and background filtering as implemented by the NOMAD database (http://derisilab.ucsf.edu). Time-series sampling and experiments with synchronized ex vivo culturing parasites. Each experiment has treatment and controls, and starts at a specific time of post invasion. For an example of quinine treatment, the treatments start from late ring stage through trophozoite stage of the parasites. First, IC50 is determined by drug assay with synchronized parasites (5% parasiteomia and 2% RBC). Second, synchronized parasite cultures are splitted into 12 flasks (75ml culture/flask) for a 6 time-point time-series experiment. 6 flasks are treated with quinine (final concentration is IC50) and 6 flasks are negative controls. 1,2,4,6,8 and 10 hrs after the treatment, parasites in each flask were harvested for total RNA isolation and microarray hybridizaiton.

2010-05-18 | E-GEOD-19468 | biostudies-arrayexpress

Heterochromatin protein 1 secures survival and transmission of malaria parasites

Project description:The malaria parasite Plasmodium falciparum relies on clonally variant gene expression in order to escape immune recognition and secure continuous proliferation during blood stage infection. Here, we studied the role of heterochromatin protein 1 (HP1), an evolutionary conserved regulator of heritable gene silencing, in the biology of P. falciparum blood stage parasites. We demonstrate that conditional PfHP1 depletion de-represses hundreds of heterochromatic virulence genes and disrupts the elusive mechanism underlying mutually exclusive expression and antigenic variation of PfEMP1. Intriguingly, we also discovered that the PfHP1-dependent regulation of an ApiAP2 transcription factor controls the switch from asexual parasite proliferation to sexual differentiation. This uncovers the first mechanistic insight into the unknown pathway triggering gametocyte conversion and establishes a new concept of HP1-dependent cell fate decision in unicellular eukaryotes. P. falciparum 3D7 parasites expressing endogenous PfHP1-GFP-DD were grown in presence of 4nM WR/625nM Shield-1 (3D7/HP1ON) or 4nM WR (3D7/HP1OFF). RNA extracted from these samples at eleven consecutive time points each was processed for microarray analysis.

2014-07-09 | E-GEOD-53176 | biostudies-arrayexpress

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2016-06-01 | E-GEOD-72579 | biostudies-arrayexpress

Artemisinin resistance in Plasmodium falciparum is associated with an altered temporal pattern of transcription (CGH)

Project description:Artemisinin resistance in Plasmodium falciparum malaria has emerged in western Cambodia. This is a major threat to global plans to control and eliminate malaria as the artemisinins are a key component of antimalarial treatment throughout the world. Using DNA microarrays we identify key features of a transcriptional profile that are associated with the delayed parasite clearance phenotype. These include reduced expression of several basic metabolic and cellular pathways in the early stages, and increased expression of essentially all functionalities associated with protein metabolism in the later stages of P. falciparum intraerythrocytic development. This is consistent with the reduced ring stage susceptibility that characterizes artemisinin resistant P. falciparum. This modulation of the P. falciparum intraerythrocytic transcriptome may result from differential expression of several regulatory proteins such as transcription factors of chromatin remodeling associated factors. In addition, the artemisinin resistant phenotype is strongly associated with a specific pattern of copy number variations, some of which are linked with differential expression of several regulatory proteins such as histone 4 and zinc permease. This study reports the first global transcriptional survey of artemisinin resistant parasites and provides a set of candidate genes for further investigation. 6 P. falciparum parasites (field isolates) which are either Artemsinin resistant or sensitive from 3 study sites (Pailin in Cambodia, Xepon in Laos, Mae Sot in Thailand) were sampled and harvested for genomic DNA. gDNA from a total of 6 samples were extracted by phenol chloroform. Synthesis of labelled target DNA was carried out as previously described: Mackinnon, M.J. et al. Comparative transcriptional and genomic analysis of Plasmodium falciparum field isolates. PLoS Pathog 5, e1000644 (2009), and used in comparative genomic microarray hybridizations (CGH).

2010-12-07 | E-GEOD-25879 | biostudies-arrayexpress

Mutually Exclusive Transcription of Subtelomeric Gene Families in Plasmodium falciparum is Restricted to var Genes

Project description:The P. falciparum genome is equipped with several subtelomeric gene families that are implicated in parasite virulence and immune evasion. The members of these gene families are uniformly positioned within heterochromatic domains of the genome and are thus subject to variegated expression. The best-studied example is that of the var gene family encoding the major parasite virulence factor P. falciparum erythrocyte membrane protein 1 (PfEMP1). Transcriptional regulation of other subtelomeric gene families and their role in parasite biology is much less understood. Here, we investigated the mode of transcriptional control of var, rif, stevor, phist and pfmc-2tm families by comparative genome-wide transcriptional profiling of transgenic parasite lines. Our results establish a clear functional distinction between var and non-var transcriptional control mechanisms. Unlike var promoters, we find that promoters of non-var families are not silenced by default. Moreover, we show that mutually exclusive transcription is unique to the var gene family. 3D7 wild-type parasites were transfected with constructs carrying eight different promoters that drive expression of the drug-selectable marker hdhfr-gfp. Thereof seven promoters are members of the multigene families upsA var, upsB var, upsC var, rif, stevor, phistb and pfmc-2tm. The cam promoter was used as transfection-based control and also a wild-type 3D7 cell line was included as control. These nine cell lines were subjected to genome-wide transcriptional profiling. Parasites were synchronized to obtain an 8 hour growth window and were harvested at four consecutive timepoints (TP): TP1 (6-14 hours post-invasion (hpi)); TP2 (14-22 hpi); TP3 (22-30 hpi); TP4 (30-38 hpi) to monitor intra- and inter-family specific linkage of multigene family expression.

2011-09-01 | E-GEOD-31829 | biostudies-arrayexpress

Artemisinin resistance in Plasmodium falciparum is associated with an altered temporal pattern of transcription (expression)

Project description:Artemisinin resistance in Plasmodium falciparum malaria has emerged in western Cambodia. This is a major threat to global plans to control and eliminate malaria as the artemisinins are a key component of antimalarial treatment throughout the world. Using DNA microarrays we identify key features of a transcriptional profile that are associated with the delayed parasite clearance phenotype. These include reduced expression of several basic metabolic and cellular pathways in the early stages, and increased expression of essentially all functionalities associated with protein metabolism in the later stages of P. falciparum intraerythrocytic development. This is consistent with the reduced ring stage susceptibility that characterizes artemisinin resistant P. falciparum. This modulation of the P. falciparum intraerythrocytic transcriptome may result from differential expression of several regulatory proteins such as transcription factors of chromatin remodeling associated factors. In addition, the artemisinin resistant phenotype is strongly associated with a specific pattern of copy number variations, some of which are linked with differential expression of several regulatory proteins such as histone 4 and zinc permease. This study reports the first global transcriptional survey of artemisinin resistant parasites and provides a set of candidate genes for further investigation. 11 P. falciparum parasites (field isolates) which are either Artemsinin resistant or sensitive from 3 study sites (Pailin in Cambodia, Xepon in Laos, Mae Sot in Thailand) were sampled, grown ex-vivo over 48 hours and harvested at regular intervals. RNA from a total of 91 samples were extracted. Synthesis of target DNA was carried out as previously described: Mackinnon, M.J. et al. Comparative transcriptional and genomic analysis of Plasmodium falciparum field isolates. PLoS Pathog 5, e1000644 (2009), and used in microarray hybridizations.

2010-12-07 | E-GEOD-25878 | biostudies-arrayexpress

Dynamics of epigenetic regulation of gene expression during Plasmodium falciparum life cycle

Project description:In order to study the role of chromatin remodeling in transcriptional regulation associated with the progression of the P. falciparum intraerythrocytic development cycle (IDC), we mapped the temporal pattern of chromosomal association with histone H3 and H4 modifications using chromatin-immunoprecipitation coupled to microarray (ChIP-on-chip). Genome-wide distribution of 13 histone modifications for 6 h time points of the 48 h P. falciparum IDC was done using ChIP-on-chip and compared to corresponding transcriptional profiles across the genome

2013-03-02 | E-GEOD-39238 | biostudies-arrayexpress

P. falciparum (lab strain DD2) treated with ionomycin vs untreated [CGH]

Project description:Calcium is a universal second messenger molecule which plays a significant role in several biological processes. Presence of calcium sensors (calmodulins) and calcium-dependent protein kinases in Plasmodium species suggests an important role of calcium-dependent signaling pathways in the regulation of cellular processes in the malaria parasites. Evidence for the transcriptional response of Plasmodium falciparum asexual blood stages to the well-known calcium ionophore ionomycin has been presented here. Comparative genomic hybridization (CGH) was carried out with total DNA isolated from the untreated parasites and the parasites treated with the calcium ionophores (ionomycin 5 Î¼M concentration) at 1 hour, 4 hours and 6 hours post treatment. Genomic DNA from the isolates was extracted by phenol chloroform. 3 Î¼g of the total DNA from each of the isolate was subjected to klenow(NEB) reaction as described in Bozdech Z, Llinas M, Pulliam BL, Wong ED, Zhu J, DeRisi JL: The transcriptome of the intraerythrocytic developmental cycle of Plasmodium falciparum. PLoS Biol 2003, 1(1):E5. Treated DNA labeled with Cy3 was then hybridized against untreated DNA labeled with Cy5.

2012-07-14 | E-GEOD-33605 | biostudies-arrayexpress

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