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Transgenic Plasmodium falciparum parasites line 3D7/DDGFP-PfAP2-HC: transcriptional differences between 3D7/DDGFP-PfAP2-HC control parasites (cultured in presence of Shield-1; ON) vs PfAP2-HC-depleted parasites (cultured in absence of Shield-1; OFF)

ABSTRACT: Transcriptional profiling of transgenic P. falciparum asexual blood stage parasites of the transgenic strain 3D7/DDGFP-PfAP2-HC at five time points during intra-erythrocytic parasite development. The DD (FKBP destabilisation domain) allows for the conditional expression of fusion proteins: DD fusion proteins are rapidly degraded or stably expressed in absence or presence of the stabilising ligand Shield-1, respectively (Banaszynski LA, Chen LC, Maynard-Smith LA, Ooi AG, Wandless TJ. A rapid, reversible, and tunable method to regulate protein function in living cells using synthetic small molecules.Cell. 2006 Sep 8;126(5):995-1004). The goal of this experiment was to identify genes differentially expressed in DDGFP-PfAP2-HC-expressing compared to DDGFP-PfAP2-HC-depleted parasites during the intra-erythtrocytic cell cycle.

ORGANISM(S): Plasmodium falciparum

PROVIDER: GSE159061 | GEO | 2020/10/06

REPOSITORIES: GEO

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Transgenic Plasmodium falciparum parasites line 3D7/DDGFP-PfAP2-HC: transcriptional differences between 3D7/DDGFP-PfAP2-HC control parasites (cultured in presence of Shield-1 ON) vs PfAP2-HC-depleted parasites (cultured in absence of Shield-1 OFF)

Project description:Transgenic Plasmodium falciparum parasites line 3D7/DDGFP-PfAP2-HC: transcriptional differences between 3D7/DDGFP-PfAP2-HC control parasites (cultured in presence of Shield-1 ON) vs PfAP2-HC-depleted parasites (cultured in absence of Shield-1 OFF)

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PfAP2-HC, an unusual, heterochromatin-associated ApiAP2 factor of Plasmodium falciparum

Project description:In this study we have investigated PfAP2-HC (PF3D7_1456000), a protein that was identified by co-immunoprecipitation with PfHP1 coupled with liquid chromatography-tandem mass spectrometry. PfAP2-HC belongs to the ApiAP2 family, the main transcription factor family in Apicomplexan parasites. We have confirmed that AP2-HC colocalises with HP1 with the use of immunofluorescence assays and chromatin immunoprecipitation-sequencing. We show that PfAP2-HC is not required for heterochromatin maintenance and inheritance with the use of PfAP2-HC Kock out and knockdown. We show with transcriptome-wide microarray time course analysis that PfAP2-HC does not act as a transcription factor in blood stage parasites. We demonstrate that the AP2 domain is dispensable for heterochromatin targeting by introducing a premature stop codon before the AP2 domain. We show that PfAP2-HC binding to heterochromatin dependents on PfHP1. and PfAP2-HC is likely not involved in de novo heterochromatin formation

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Transgenic Plasmodium falciparum blood stage parasites 3D7/GDV1-GFP-DD: control (-Shield-1;3D7/GDV1-GFP-DDOFF) vs GDV1-overexpressing (+Shield-1;3D7/GDV1-GFP-DDON) parasites

Project description:Transcriptional profiling of transgenic P. falciparum blood stage parasites expressing an ectopic copy of GDV1-GFP-DD (3D7/GDV1-GFP-DD). The DD (FKBP destabilisation domain) allows for the conditional expression of fusion proteins: DD fusion proteins are rapidly degraded or stably expressed in absence or presence of the stabilising ligand Shield-1, respectively (Banaszynski LA, Chen LC, Maynard-Smith LA, Ooi AG, Wandless TJ. A rapid, reversible, and tunable method to regulate protein function in living cells using synthetic small molecules.Cell. 2006 Sep 8;126(5):995-1004). The goal of this experiment was to identify genes differentially expressed in response to induced GDV1 overexpression.

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Transgenic Plasmodium falciparum blood stage parasites F12/GDV1-GFP-DD: control (-Shield-1;F12/GDV1-GFP-DDOFF) vs GDV1-overexpressing (+Shield-1;F12/GDV1-GFP-DDON) parasites

Project description:Transcriptional profiling of transgenic P. falciparum blood stage parasites expressing an ectopic copy of GDV1-GFP-DD (F12/GDV1-GFP-DD). The DD (FKBP destabilisation domain) allows for the conditional expression of fusion proteins: DD fusion proteins are rapidly degraded or stably expressed in absence or presence of the stabilising ligand Shield-1, respectively (Banaszynski LA, Chen LC, Maynard-Smith LA, Ooi AG, Wandless TJ. A rapid, reversible, and tunable method to regulate protein function in living cells using synthetic small molecules.Cell. 2006 Sep 8;126(5):995-1004). The goal of this experiment was to identify genes differentially expressed in response to induced GDV1 overexpression.

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PfAP2-I-GFP-msp5MUT vs PfAP2-I-GFP 3D7 [timecourse 2]

Project description:Obligate intracellular parasites must efficiently invade host cells in order to mature and be transmitted. For the malaria parasite Plasmodium falciparum, invasion of host red blood cells (RBCs) is essential. Here we describe a parasite-specific transcription factor belonging to the Apicomplexan Apetala 2 (ApiAP2) family that is responsible for regulating the expression of a subset of merozoite genes involved in RBC invasion (PfAP2-I). Our genome-wide analysis by ChIP-seq shows that PfAP2-I interacts with a specific DNA motif in the promoters of these genes. msp5 transcription levels decrease when the PfAP2-I DNA-binding motif is mutated in PfAP2-I-GFP parasites, showing that PfAP2-I must bind the DNA motif in order for msp5 to be transcribed.

2016-12-31 | GSE77809 | GEO

PfAP2-I-GFP-msp5MUT vs PfAP2-I-GFP 3D7 [timecourse 1]

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A transcriptional repressor programs gametocytogenesis in human malaria parasites (PfAP2-G5) [WGS-Seq]

Project description:Gametocytogenesis, the process by which malaria parasites produce sexual forms that can infect mosquitoes, is essential for the transmission of malaria. A transcriptional switch of the pfap2-g gene triggers sexual commitment, but how the complex multi-step process is precisely programed remains largely unknown. Here, by systematic functional screening of a panel of ApiAP2 transcription factors, we identify six new apiAP2 members associated with gametocytogenesis in Plasmodium falciparum. Among these, PfAP2-G5 (PF3D7_1139300) was found to be indispensable for gametocytogenesis. This factor suppresses the transcriptional activity of the pfap2-g gene via binding to both the upstream region and exonic gene body, the latter is linked to the maintenance of local heterochromatin structure, thereby preventing initiation of sexual commitment. Removal of this repressive effect through pfap2-g5 knockout disrupts the asexual replication cycle and promotes sexual commitment accompanied by upregulation of pfap2-g expression. However, the gametocytes produced fail to mature fully. Further analyses show that PfAP2-G5 is essential for gametocyte maturation, and causes the down-regulation of pfap2-g and a set of early gametocyte genes activated by PfAP2-G prior to gametocyte development. Collectively, our findings reveal a regulation cascade of gametocyte production in malaria parasites, and provide a new target for transmission blocking interventions.

2021-08-03 | GSE175931 | GEO

A transcriptional repressor programs gametocytogenesis in human malaria parasites (PfAP2-G5) [RNA-Seq]

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A transcriptional repressor programs gametocytogenesis in human malaria parasites (PfAP2-G5) [ChIP-Seq HM]

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A transcriptional repressor programs gametocytogenesis in human malaria parasites (PfAP2-G5) [ChIP-Seq TF]

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