Plasmodium falciparum whole-genome real-time transcription and decay
Ontology highlight
ABSTRACT:
ORGANISM(S): Plasmodium falciparum
PROVIDER: GSE66669 | GEO | 2015/10/01
SECONDARY ACCESSION(S): PRJNA277701
REPOSITORIES: GEO
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Project description:To date, total mRNA analysis throughout intraerythrocytic development of the malaria parasite, Plasmodium falciparum, has only revealed abundance profiles of each gene at a given time. Here, we establish a new methodology in Plasmodium falciparum that enables biosynthetic labeleing and capture of sub-population mRNA. As a proof of principle for this novel method, we examine the mRNA dynamics of early gametocyte commitment.
2015-09-04 | GSE72695 | GEO
Project description:The rate of mRNA decay is an essential element of post-transcriptional regulation in all organisms. Previously, studies in several organisms found that the specific half-life of each mRNA is precisely related to its physiological role, and plays an important role in determining levels of gene expression. We have used a genome wide approach to characterize mRNA decay in Plasmodium falciparum. We found that globally, rates of mRNA decay increase dramatically during the asexual intraerythrocytic developmental cycle. During the ring stage of the cycle, the average mRNA half-life was 9.5 minutes, yet this was extended to an average of 65 minutes during the late schizont stage of development. Thus a major determinant of mRNA decay rate appears to be linked to the stage of intraerythrocytic development. Furthermore, we have found specific variations in decay patterns superimposed upon the dominant trend of progressive half-life lengthening. These variations in decay pattern were frequently enriched for genes with specific cellular functions or processes. Elucidation of Plasmodium mRNA decay rates provides a key element for deciphering mechanisms of genetic control in this parasite, by complementing and extending previous mRNA abundance studies. Our results indicate that progressive stage-dependent decreases in mRNA decay rate function are a major determinant of mRNA accumulation during the schizont stage of intraerythrocytic development. This type of genome wide change in mRNA decay rate has not been observed in any other organism to date, and indicates that post-transcriptional regulation may be the dominant mechanism of gene regulation in P. falciparum. Keywords: Plasmodium falciparum treated with actinomycin D
2007-07-19 | GSE8099 | GEO
Project description:It is widely assumed that sex-determination loci have little impact upon dominant stage in the life history of filamentous fungi, asexual development. However, this hypothesis has not been thoroughly investigated. To investigate differences between mating types during asexual development, total RNA was sampled from otherwise largely isogenic Mat-A and Mat-a N. crassa strains at early, middle, and late clonal stages of development under a condition of constant light. Mating-type loci, pheromone precursor and receptor genes were assayed with real-time PCR. Mating loci were increasingly expressed during asexual development, and expression of pheromone precursors ccg-4 and mfa-1 and receptors pre-1 and pre-2 were detected in both mating types in all development stages. A circuit of microarray hybridizations of cDNA samples from all developmental stages was also performed. Gene expression for both mating types throughout vegetative development were characterized for developmental markers such as transcription factors, for genes related to conidiation, internal clock, cell division cycle, heat shock, and for light responsive genes. We observed significant differences in overall gene expression between the mating types across clonal development, especially at late development stages. The Mat-A genotype showed a higher expression level than Mat-a for many genes, and demonstrated greater transcriptional regulatory activity. Significant up-regulation of expression was observed for many late light responsive genes at late asexual development stages in both mating types. Further investigation of the impact of light and the roles of light response genes in asexual development in this fungus are warranted. Neurospora crassa mat A FGSC#2489 and mat-a FGSC 4200: Three developmental stages, 36hr, 60hr, 96 hr were sampled and compared
2011-06-06 | E-GEOD-26209 | biostudies-arrayexpress
Project description:It is widely assumed that sex-determination loci have little impact upon dominant stage in the life history of filamentous fungi, asexual development. However, this hypothesis has not been thoroughly investigated. To investigate differences between mating types during asexual development, total RNA was sampled from otherwise largely isogenic Mat-A and Mat-a N. crassa strains at early, middle, and late clonal stages of development under a condition of constant light. Mating-type loci, pheromone precursor and receptor genes were assayed with real-time PCR. Mating loci were increasingly expressed during asexual development, and expression of pheromone precursors ccg-4 and mfa-1 and receptors pre-1 and pre-2 were detected in both mating types in all development stages. A circuit of microarray hybridizations of cDNA samples from all developmental stages was also performed. Gene expression for both mating types throughout vegetative development were characterized for developmental markers such as transcription factors, for genes related to conidiation, internal clock, cell division cycle, heat shock, and for light responsive genes. We observed significant differences in overall gene expression between the mating types across clonal development, especially at late development stages. The Mat-A genotype showed a higher expression level than Mat-a for many genes, and demonstrated greater transcriptional regulatory activity. Significant up-regulation of expression was observed for many late light responsive genes at late asexual development stages in both mating types. Further investigation of the impact of light and the roles of light response genes in asexual development in this fungus are warranted.
2011-06-06 | GSE26209 | GEO
Project description:Genome-wide occupancy of sixteen PfApiAP2 transcription factors and PfHP1 throughout the intraerythrocytic cycle
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Project description:To date, total mRNA analysis throughout intraerythrocytic development of the malaria parasite, Plasmodium falciparum, has only revealed abundance profiles of each gene at a given time. Here, we establish a new methodology in Plasmodium falciparum that enables biosynthetic labeleing and capture of sub-population mRNA. As a proof of principle for this novel method, we examine the mRNA dynamics of early gametocyte commitment. Plasmodium falciparum strains 3D7, E5, and F12 were highly synchronized and, at 36 hours post invasion, incubated with 40um 4-TU for 12 hours followed immediately by Trizol total RNA extraction. Total RNA from each timepoint was then biotinylated via a thiol-group on any transcript that incorporated a thiol-modified UTP. Biotinylated transcripts were then separated from total RNA by Streptavidin magnetic beads resulting in a Labeled sample. Any mRNA that was not bound to the beads resulted in an Unlabeled sample. Every 12 hours, two samples were analyzed by Agilent P. falciparum DNA microarray, Unlabeled and Labeled, resulting in 48 individual DNA microarrays (4 for each sample type).
2015-09-04 | E-GEOD-72695 | biostudies-arrayexpress
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 control Plasmodium falciparum asexual blood stages not treated with the calcium ionophores, A23187 and ionomycin has been presented here. P. falciparum 3D7 strain was cultured as described by 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. Total RNA from each of the time points was isolated and aminoallyl-cDNA was synthesized using reverse transcriptase system (Fermentas). cDNA made from the untreated parasites were labeled with Cy5 (GE-Amersham). A reference pool was made by mixing equal amount of cDNA from the parasites collected at 6 hours interval throughout the 48 hours life cycle and was labeled with Cy3 (GE-Amersham). The samples were then hybridized on a spotted cDNA chip platform comprising 10166 MOEs representing 5363 coding sequences as described in Hu G, Cabrera A, Kono M, Mok S, Chaal BK, Haase S, Engelberg K, Cheemadan S, Spielmann T, Preiser PR, Gilberger TW, Bozdech Z: Transcriptional profiling of growth perturbations of the human malaria parasite Plasmodium falciparum. Nat Biotechnol, 2009. 28(1): p. 91-8. The data was normalized and filtered with the condition, signal intensity>background intensity + 2 SD of background intensity) using NOMAD.
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Project description:T. gondii has a complex life cycle typified by an asexual development taking place in vertebrate, and a sexual reproduction which occurs exclusively in felids and thereby is less studied. The developmental transitions rely on changes in gene expression patterns, and recent studies have assigned roles for chromatin shapers, including histone modifications, in establishing specific epigenetic programs for each given stage. Here, we identified T. gondii microrchidia (MORC) protein as an upstream transcriptional repressor of sexual commitment. We performed affinity purification coupled to tandem mass spectrometry analyses to identify its binding partners.
2020-01-09 | PXD016846 | Pride
Project description:Histone variants are key components of the epigenetic code and evolved to perform specific functions in transcriptional regulation, DNA repair, chromosome segregation and other fundamental processes. H2B.Z is a rare, apicomplexan-specific variant of histone H2B. Here we show that in Plasmodium falciparum H2B.Z localises to euchromatic intergenic regions throughout intraerythrocytic development and together with H2A.Z forms a double-variant nucleosomes subtype. These nucleosomes are enriched in promoters over 3’ intergenic regions and their occupancy generally correlates with the strength of the promoter, but not with its temporal activity. Remarkably, H2B.Z occupancy levels exhibit a clear correlation with the base-composition of the underlying DNA, raising the intriguing possibility that the extreme AT-content of the intergenic regions within the Plasmodium genome might be instructive for histone variant deposition. In summary, our data shows that the H2A.Z/H2B.Z double-variant nucleosome demarcates putative regulatory regions of the P. falciparum epigenome and likely provides a scaffold for dynamic regulation of gene expression in this deadly human pathogen. Genome-wide localization of H2B.Z has been studied at three stages of intraerythrocytic development by Illumina sequencing of chromatin-immunoprecipitated and input DNA.
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Project description:In eukaryotes, the chromatin architecture has a pivotal role in regulating all DNA-related processes. For P. falciparum, the causative agent of human malaria, the nucleosome landscape of the extremely AT-rich intergenic regulatory regions is largely unexplored. With the aid of a highly controlled MNase-seq procedure we reveal how positioning of nucleosomes provides a structural and regulatory framework to the transcriptional unit. We observe strong positioning of nucleosomes around splice sites that could aid co-transcriptional splicing events. In addition, nucleosome depleted regions are apparent hallmarks of transcription start sites (TSSs) and may support pre-initiation complex assembly. Moreover, we reveal nucleosome occupancy dynamics on strong TSSs during intraerythrocytic development, which correlate with gene expression changes and we observe a characteristic nucleosome architecture of functional - but not inert - TGCATGCA DNA motifs. Collectively, these findings highlight the regulatory capacity of the nucleosome landscape of this deadly human pathogen. Mnase-seq during the intra-erythrocytic asexual cycle of Plasmodium falciparum var2csa-panned 3D7 parasites for 8 time-points, every 5 hours starting from 5 hours post invasion until 40 hours post-invasion (T5-T40). Cycle length of these parasites is ~43 hours, synchronicity window is ~ 8 hours. T40 has 2 technical replicates (independent digestions; T40A, T40B). Additionally, pellet control sample (T15), histone H4-ChIP control (T40A) and sonicated and amplified genomic DNA. Chromatin was digested using a combined MNase + exonuclease III treatment. Libraries were prepared according to a Plasmodium-optimized library preparation procedure including KAPA polymerase-mediated PCR amplification. Strand-specific RNA-seq for expression quantification during the intra-erythrocytic asexual cycle of Plasmodium falciparum var2csa-panned 3D7 parasites for 8 time-points every 5 hours starting from 5 hours post invasion invasion until 40 hours post-invasion (T5-T40). Cycle length of these parasites is ~43 hours, synchronicity window is ~ 8 hours. These samples are originating from the exact same batch of parasites as are the MNase-Seq libraries. Libraries were prepared according to a Plasmodium-optimized library preparation procedure including KAPA polymerase-mediated PCR amplification.
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