Coordinated regulation of gene expression in Plasmodium female gametocytes by two transcription factors
- Department of Medical Zoology, Mie University School of Medicine, Japan
- Department of Molecular Protozoology, Research Center for Infectious Disease Control, Japan
Figures
Figure 1 with 1 supplement
PFG is a target gene of AP2-G and is expressed in female gametocytes.
(A) Schematic diagram of PFG from P. berghei and its homologs in apicomplexan parasites. Regions homologous to regions 1 and 2, which are highly conserved among Plasmodium species, are shown as …
Figure 1—figure supplement 1
Preparation of PFG::GFP parasites.
-
Figure 1—figure supplement 1—source data 1
Original gel image for Figure 1—figure supplement 1.
- https://cdn.elifesciences.org/articles/88317/elife-88317-fig1-figsupp1-data1-v1.pdf
Figure 2 with 1 supplement
Development of female gametocytes was impaired in PFG(-)parasites.
(A) Proliferation of blood-stage parasites in wild-type and PFG(−) parasites. Parasitemia was calculated by counting infected red blood cells using Giemsa staining of blood smears. Three …
Figure 2—figure supplement 1
Preparation of PFG(-) and PFG(-)820 parasites.
-
Figure 2—figure supplement 1—source data 1
Original gel image for Figure 2—figure supplement 1.
- https://cdn.elifesciences.org/articles/88317/elife-88317-fig2-figsupp1-data1-v1.pdf
Figure 3
PFG is co-localized with AP2-FG.
(A) IGV images from ChIP-seq experiments 1 and 2 of PFG on chromosome 14. Histograms show row read coverage in the ChIP data at each base. The Venn diagram on the right shows the number of peaks …
Figure 4 with 2 supplements
PFG mediates AP2-FG binding to the ten-base motif.
(A) Venn diagram showing the number of common peaks between experiments 1 and 2 in ChIP-seq of PFG using PFG::GFPAP2-FG(-) parasites. Peaks were regarded as common when their summits were within …
Figure 4—figure supplement 1
Preparation of PFG::GFPAP2-FG(-) parasites.
PFG::GFPAP2-FG(-) parasites were prepared from AP2-FG(-) parasites using the sulfadiazine-resistant dihydropteroate synthase (DHPS) gene as a selectable marker gene.
-
Figure 4—figure supplement 1—source data 1
Original gel image for Figure 4—figure supplement 1.
- https://cdn.elifesciences.org/articles/88317/elife-88317-fig4-figsupp1-data1-v1.pdf
Figure 4—figure supplement 2
Preparation of AP2-FG::GFPPFG(-) parasites.
AP2-FG::GFPPFG(-) parasites were prepared using CRISPR/Cas9 in two steps. First, PFG was disrupted in pbcas9, and then GFP was fused to the 3’-portion of the AP2-FG gene.
-
Figure 4—figure supplement 2—source data 1
Original gel image for Figure 4—figure supplement 2.
- https://cdn.elifesciences.org/articles/88317/elife-88317-fig4-figsupp2-data1-v1.pdf
Figure 5
sAP2-FG binds to five-base motifs that function as a cis-acting element independent of the ten-base motif.
(A) Motifs enriched around peaks identified in DIP-seq of the AP2 domain of AP2-FG. Sequence logos are depicted using WebLogo 3. (B) Histogram showing the distance between the peak summit and the …
Figure 6
Target genes of PFG (cAP2-FG) and sAP2-FG contain different sets of genes.
(A) Functional classification of the PFG target genes (cAP2-FG) (572 annotated genes in total). In this graph, different subgroups related to female gametocyte-specific functions are collectively …
Author response image 1
Legged: The Venn diagram shows the number of common peaks between these ChIP seq experiments (distance of peak summits < 150).
Author response image 2
Additional files
-
Supplementary file 1
Differential expression analysis between wild-type and PFG(-) parasites.
- https://cdn.elifesciences.org/articles/88317/elife-88317-supp1-v1.xlsx
-
Supplementary file 2
Peaks identified in ChIP-seq experiments and their nearest genes.
(a) Peaks identified in ChIP-seq of PFG with their nearest genes (experiment 1). (b) Peaks identified in ChIP-seq of PFG (experiment 2). (c) Peaks identified in ChIP-seq of PFG using AP2-FG(-) parasites (experiment 1). (d) Peaks identified in ChIP-seq of PFG using AP2-FG(-) parasites (experiment 2). (e) Peaks identified in ChIP-seq of sAP2-FG (experiment 1). (f) Peaks identified in ChIP-seq of sAP2-FG (experiment 2). (g) Peaks identified in DIP-seq using recombinant AP2 domain of AP2-FG and P. berghei genomic DNA.
- https://cdn.elifesciences.org/articles/88317/elife-88317-supp2-v1.xlsx
-
Supplementary file 3
Target genes predicted from ChIP-seq peaks.
(a) PFG target genes predicted from ChIP-seq peaks. (b) sAP2-FG target genes predicted from ChIP-seq peaks.
- https://cdn.elifesciences.org/articles/88317/elife-88317-supp3-v1.xlsx
-
Supplementary file 4
List of primers used in this study.
- https://cdn.elifesciences.org/articles/88317/elife-88317-supp4-v1.xlsx
-
MDAR checklist
- https://cdn.elifesciences.org/articles/88317/elife-88317-mdarchecklist1-v1.docx
