RNA polymerase III is involved in regulating Plasmodium falciparum virulence
- Institut Pasteur, Universite Paris Cité, France
- Institut Pasteur, Sorbonne Université Ecole doctorale Complexité du Vivant, France
- Institut Pasteur, Biomedical Center, Division of Physiological Chemistry, Faculty of Medicine, Ludwig-Maximilians-Universität München, Germany
- Institut Pasteur, Laboratory of Pathogen-Host Interaction (LPHI), CNRS, University of Montpellier, France
- Institut Pasteur, Institut Curie, PSL Research University, Centre de Recherche, CurieCoreTech Mass Spectrometry Proteomics, France
- Institut Pasteur, Instituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina da Universidade de Lisboa, Portugal
- Institut Pasteur, LPHI, MIVEGEC, CNRS, INSERM, University of Montpellier, France
Figures
Figure 1 with 2 supplements
RNA polymerase III (RNA Pol III)-transcribed genes are downregulated in asymptomatic individuals during the dry season.
Steady-state RNA levels as quantified by reverse transcription-quantitative PCR (RT-qPCR) using primers to tRNA Asparagine (Pf3D7_0714700) (A) and tRNA Valine (Pf3D7_0312600) (B) as well as RNA Pol …
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Figure 1—source data 1
Reverse transcription-quantitative PCR (RT-qPCR) values used in analysis.
- https://cdn.elifesciences.org/articles/95879/elife-95879-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
RUF6 is transcribed by RNA polymerase III (RNA Pol III).
(A) Multiple sequence alignment of highly conserved genes encoding GC-rich ncRNA elements. 15 members of RUF6 gene family aligned by Clustal Omega (http://www.ebi.ac.uk) and presented in Jalview (htt…
Figure 1—figure supplement 2
RNA polymerase III (RNA Pol III)-transcribed genes are reduced during dry asymptomatic cases.
(A) tRNA Leucine (Pf3D7_API05400) and tRNA Asparagine (Pf3D7_API05500) transcripts identified by RNA-seq analysis from Andrade et al., 2020, for wet symptomatic (n=12) and dry asymptomatic (n=12) …
Figure 2 with 2 supplements
External factors modulate RNA polymerase III (RNA Pol III)-transcribed genes.
(A) Plasma magnesium levels are significantly increased in asymptomatic individuals during the dry season compared to symptomatic individuals during the wet season. Concentration is shown in nmol/μL …
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Figure 2—source data 1
RNA-seq analysis for control and MgCl2 supplemented cultured parasites.
- https://cdn.elifesciences.org/articles/95879/elife-95879-fig2-data1-v1.xlsx
Figure 2—figure supplement 1
RNA polymerase III (RNA Pol III)-transcribed genes are reduced in response to external factors.
Transcript levels as quantified by reverse transcription-quantitative PCR (RT-qPCR) on 3D7 parasites harvested during late ring stage for control parasites, parasites in the absence of isoleucine …
Figure 2—figure supplement 2
RNA polymerase III (RNA Pol III)-transcribed genes are reduced in response to increasing MgCl2 supplementation concentrations.
(A) Transcript levels as quantified by reverse transcription-quantitative PCR (RT-qPCR) on 3D7 parasites harvested during late ring stage for control parasites and parasites in the presence of …
Figure 3 with 1 supplement
Nuclear PfMaf1 is essential to regulate RNA polymerase III (RNA Pol III).
(A) Illustration of the TORC1-dependent cellular localization of Maf1 protein in unfavorable conditions in typical eukaryotic organisms compared to P. falciparum that has no TORC1 pathway. (B, top) …
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Figure 3—source data 1
Western blot analysis shows PfMaf1 is depleted after 1 and 3 cycles without Shield-1 drug added.
- https://cdn.elifesciences.org/articles/95879/elife-95879-fig3-data1-v1.pdf
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Figure 3—source data 2
Western blot analysis shows PfMaf1 is depleted after 2 cycles without Shield-1 drug added.
- https://cdn.elifesciences.org/articles/95879/elife-95879-fig3-data2-v1.pdf
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Figure 3—source data 3
Co-immunoprecipitation followed by quantitative mass spectrometry (Co-IP MS) analysis for cytoplasmic and nuclear PfMaf1-binding proteins.
- https://cdn.elifesciences.org/articles/95879/elife-95879-fig3-data3-v1.xls
Figure 3—figure supplement 1
PfMaf1 cytoplasmic and nuclear interactome.
Co-immunoprecipitation followed by quantitative mass spectrometry (Co-IP MS) volcano plot of enrichment for all five replicates for cytoplasmic (A) and nuclear (B) PfMaf1 vs control proteins are …
Figure 4 with 1 supplement
External factors modulate virulence through PfMaf1-regulated RNA polymerase III (RNA Pol III) inhibition.
(A) Immunoprecipitation western blot analysis for cytoplasmic and nuclear extracts for PfMaf1 expression in PfMaf1-FKBP transfected parasites with addition of MgCl2 ([3 mM] total) and control …
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Figure 4—source data 1
Immunoprecipitation followed by western blot analysis shows nuclear PfMaf1 levels increase with MgCl2 supplementation.
- https://cdn.elifesciences.org/articles/95879/elife-95879-fig4-data1-v1.pdf
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Figure 4—source data 2
Western blot analysis shows decreased PfEMP1 expression with MgCl2 supplementation.
- https://cdn.elifesciences.org/articles/95879/elife-95879-fig4-data2-v1.pdf
Figure 4—figure supplement 1
MgCl2 supplementation increases nuclear PfMaf1 levels.
(A) Immunoprecipitation western blot analysis for cytoplasmic and nuclear extracts for PfMaf1 expression in PfMaf1-FKBP transfected parasites with addition of MgCl2 ([3 mM] total) and control …
Additional files
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Supplementary file 1
Quantitative PCR (qPCR) analysis primer pairs.
Primer pairs used are listed including the forward and reverse sequences.
- https://cdn.elifesciences.org/articles/95879/elife-95879-supp1-v1.docx
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Supplementary file 2
Samples used for plasma magnesium concentration assay.
Age and sex are indicated for each sample as well as the month each sample was from. Green highlight shows magnesium concentration and purple highlight shows which samples were also used for reverse transcription-quantitative PCR (RT-qPCR) analysis in Figure 1.
- https://cdn.elifesciences.org/articles/95879/elife-95879-supp2-v1.xlsx
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MDAR checklist
- https://cdn.elifesciences.org/articles/95879/elife-95879-mdarchecklist1-v1.pdf
