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Status |
Public on Jul 17, 2019 |
Title |
PfAP2-G ChIP-seq in stage I gametocytes derived via next cycle conversion (NCC) and same cycle conversion (SCC) |
Organism |
Plasmodium falciparum |
Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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Summary |
In the malaria parasite Plasmodium falciparum, the switch from asexual multiplication to sexual differentiation into gametocytes is essential for transmission to mosquitos. One of the key determinants of sexual commitment is the transcription factor PfAP2-G, which has been proposed to orchestrate this crucial cell fate decision by driving expression of gametocyte genes. Here we identify for the first time the direct targets of PfAP2-G and demonstrate that it dynamically binds hundreds of sites across the genome. We also identify major differences in PfAP2-G occupancy between gametocytes derived via next-cycle and same-cycle conversion, suggesting significant biological differences between these two pathways. By using a combination of ChIP-seq, transcriptomics, and CRISPR-Cas9, we are able to show definitively that PfAP2-G is a transcriptional activator of early gametocyte genes. Our data implicate PfAP2-G not just as a transcriptional activator of gametocyte genes, but also as a regulator of genes important for red blood cell invasion. Excitingly, we also demonstrate that PfAP2-G interacts with a second transcription factor to regulate transcription. These results shed new light on a critical part of the malaria parasite lifecycle by clarifying the role of PfAP2-G during gametocytogenesis.
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Overall design |
ChIP was performed using an antibody against HA on the AP2-G-DD line (+ Shld1) in stage I gametocytes produced using either next cycle conversion (NCC) or same cycle conversion (SCC). There are two biological replicates for each of the two stages, giving a total of four samples. The input material served as the control.
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Contributor(s) |
Josling GA, Russell TJ, Venezia J, Orchard L, Painter H, Llinás M |
Citation(s) |
32198457 |
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Submission date |
Jul 15, 2019 |
Last update date |
Mar 30, 2020 |
Contact name |
Manuel Llinas |
E-mail(s) |
manuel@psu.edu
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Organization name |
Pennsylvania State University
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Department |
Biochemistry and Molecular Biology
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Street address |
Millennium Science Complex
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City |
University Park |
State/province |
PA |
ZIP/Postal code |
16802 |
Country |
USA |
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Platforms (1) |
GPL26920 |
NextSeq 550 (Plasmodium falciparum) |
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Samples (8)
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Relations |
BioProject |
PRJNA554600 |
SRA |
SRP214599 |
Supplementary file |
Size |
Download |
File type/resource |
GSE134268_RAW.tar |
53.6 Mb |
(http)(custom) |
TAR (of BED, BIGWIG) |
SRA Run Selector |
Raw data are available in SRA |
Processed data provided as supplementary file |
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