Inducible developmental reprogramming redefines commitment to sexual development in the malaria parasite Plasmodium berghei

Abstract

During malaria infection, Plasmodium spp. parasites cyclically invade red blood cells and can follow two different developmental pathways. They can either replicate asexually to sustain the infection, or differentiate into gametocytes, the sexual stage that can be taken up by mosquitoes, ultimately leading to disease transmission. Despite its importance for malaria control, the process of gametocytogenesis remains poorly understood, partially due to the difficulty of generating high numbers of sexually committed parasites in laboratory conditions1. Recently, an apicomplexa-specific transcription factor (AP2-G) was identified as necessary for gametocyte production in multiple Plasmodium species2,3, and suggested to be an epigenetically regulated master switch that initiates gametocytogenesis4,5. Here we show that in a rodent malaria parasite, Plasmodium berghei, conditional overexpression of AP2-G can be used to synchronously convert the great majority of the population into fertile gametocytes. This discovery allowed us to redefine the time frame of sexual commitment, identify a number of putative AP2-G targets and chart the sequence of transcriptional changes through gametocyte development, including the observation that gender-specific transcription occurred within 6 h of induction. These data provide entry points for further detailed characterization of the key process required for malaria transmission.

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Fig. 1: Establishment of an AP2-G-overexpression system.
Fig. 2: AP2-G overexpression results in gametocyte conversion.
Fig. 3: Transcriptome changes in PBGAMiR and PBGAMiR+ parasites reveal a gametocyte-specific transcriptional programme.
Fig. 4: PBANKA_0312700 is an AP2-G-induced gene involved in gametocyte development.

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Acknowledgements

We thank R. Menard and D. Bargieri for the diCre test plasmid used in this study, and M. Sanders and the WTSI sequencing service for assistance with RNA-seq sample processing. K.K.M. is supported by the Wellcome Trust and the Royal Society (ref. 202600/Z/16/Z). R.S.K. is supported by BBSRC (ref. BB/J013854/1). A.P.W. is supported by the Wellcome Trust (refs 083811 and 107046). O.B. is supported by the Wellcome Trust Sanger Institute (ref. WT098051).

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Contributions

R.S.K. generated and phenotyped HP diCre, diCre test and PBANKA_0312700 lines, performed phenotyping experiments on the PBGAMi line and generated parasites for transcriptome sequencing. K.K.M. generated the AP2-G overexpression construct and PBGAMi line, performed phenotyping experiments on the PBGAMi line, generated RNA-seq libraries and performed RNA-seq data analysis. R.C. generated 820 diCre and GIMO lines and parasites for transcriptome sequencing. N.P. performed the ookinete conversion assay for PBANKA_0312700 line. O.B. and A.P.W. led and supervised the study. K.K.M. and A.P.W. wrote the manuscript with contributions from the other authors.

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Correspondence to Katarzyna K. Modrzynska or Oliver Billker or Andrew P. Waters.

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The authors declare no competing interests.

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Supplementary information

Supplementary Information

Supplementary Materials, Supplementary Figures 1–9, legends for Supplementary Tables 1–3 and original blots.

Reporting Summary

Supplementary Table 1

List of oligonucleotide primers used in the study.

Supplementary Table 2

Differential gene expression analysis between PBGAMiR+ and PBGAMiR populations at various time points post induction.

Supplementary Table 3

Gene ontology terms enriched in genes differentially expressed at different time points post induction.

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Kent, R.S., Modrzynska, K.K., Cameron, R. et al. Inducible developmental reprogramming redefines commitment to sexual development in the malaria parasite Plasmodium berghei. Nat Microbiol 3, 1206–1213 (2018). https://doi.org/10.1038/s41564-018-0223-6

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