Dataset Information

The translatome of quiescent Plasmodium falciparum gametocytes reveals parasite pyridoxal 5’-phosphate (PLP) biosynthesis is essential for efficient mosquito stage development

ABSTRACT: The ability of Plasmodium falciparum gametocytes to remain quiescent within the vertebrate host but poised for rapid onward development in the mosquito is an adaptation essential to maximise the onward spread of malaria. In this dormant state, mature infectious stage V gametocytes are largely unaffected by most antimalarial drugs and our limited understanding of how gametocytes prepare for mosquito transmission has hindered the identification of new molecular targets for transmission-blocking therapeutics. In this study, we move beyond the total proteome of gametocytes and define the translatome of mature stage V gametocytes using L-azidohomoalanine incorporation into nascent proteins, click chemistry purification and proteomic analysis. We identify the proteins and pathways that gametocytes sustain in preparation for transmission during this dormant period and through genetic disruption, we validate this approach by demonstrating the importance of parasite pyridoxal 5’-phosphate biosynthesis for mosquito transmission.

INSTRUMENT(S):

ORGANISM(S): Plasmodium Falciparum

DISEASE(S): Malaria

SUBMITTER: Jack Houghton

LAB HEAD: Michael Delves

PROVIDER: PXD075878 | Pride | 2026-03-25

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	JH-EAdS-WT-1_S3-B9_1_2992.d.zip	Other
	JH-EAdS-WT-2_S3-B10_1_2993.d.zip	Other
	JH-EAdS-WT-3_S3-B11_1_2994.d.zip	Other
	JH-ES-A_S4-E12_1_1450.d.zip	Other
	JH-ES-B_S4-F1_1_1451.d.zip	Other

Items per page:

1 - 5 of 11

Similar Datasets

Project description:During intra-erythrocytic development, late asexually replicating Plasmodium falciparum parasites sequester from peripheral circulation. This facilitates chronic infection and is linked to severe disease and organ-specific pathology including cerebral and placental malaria. Immature gametocytes M-bM-^@M-^S sexual stage precursor cells M-bM-^@M-^S likewise disappear from circulation. Recent work has demonstrated that these sexual stage parasites are located in the hematopoietic system of the bone marrow before mature gametocytes are released into the blood stream to facilitate mosquito transmission. However, as sequestration occurs only in vivo and not during in vitro culture, the mechanisms by which it is regulated and enacted (particularly by the gametocyte stage) remain poorly understood. We generated the most comprehensive P. falciparum functional gene network to date by integrating global transcriptional data from a large set of asexual and sexual in vitro samples, patient-derived in vivo samples, and a new set of in vitro samples profiling sexual commitment. We defined more than 250 functional modules (clusters) of genes that are co-expressed primarily during the intra-erythrocytic parasite cycle, including 35 during sexual commitment and gametocyte development. Comparing the in vivo and in vitro datasets allowed us, for the first time, to map the time point of asexual parasite sequestration in patients to 22 hours post invasion, confirming previous in vitro observations on the dynamics of host cell modification and cytoadherence. Moreover, we were able to define the properties of gametocyte sequestration, demonstrating the presence of two circulating gametocyte populations: gametocyte rings between 0 and ~30 hours post invasion and mature gametocytes after around 7 days post invasion. We used 164/TdTom, a transgenic parasite line expressing a red fluorescent protein reporter under a gametocyte-specific promoter to generate schizont samples. Schizonts were subsequently isolated from both the fluorescent and non-fluorescent population by FACS and prepared for microarray analysis. Two biological replicates were produced for both the fluorescent and the non-fluorescent samples.

Project description:Plasmodium falciparum gametocyte stages represent a small fraction of the entire parasite biomass that is present during human malaria infection, yet they alone lead to the transmission of this devastating disease. One of the critical gaps in malaria transmission biology and surveillance is our lack of knowledge about gametocyte biology, especially sexual dimorphic development that may influence transmission from the human to the mosquito. Ratios of male and female gametocytes in the peripheral blood can vary significantly; influenced in part by asexual blood stage and gametocyte density as well as vertebrate and invertebrate host factors. Moreover, the role of sex ratios on gametocyte transmission potential to mosquitoes is unknown and dissecting this process has been hampered by the lack of sex-specific protein markers for the circulating, mature stage V gametocytes. The current evidence suggests a high degree of conservation in gametocyte gene complement across Plasmodium, and therefore presumably for sex-specific genes as well. Therefore, to better our understanding of gametocyte development and subsequent infectiousness to mosquitoes, we undertook a two pronged approach. First, we acquired the mixed, male and female stage V gametocyte proteomes of the NF54 isolate and mature stage V female proteome from Dd2, a strain that is defective in producing mature males. Second, we then undertook a Systematic Subtractive Bioinformatic analysis (filtering) approach to identify sex-specific P. falciparum NF54 protein markers based on a comparison with the Dd2 strain and syntenic male and female proteins from the reanalyzed and updated P. berghei (related rodent malaria parasite) gametocyte proteomes. This has produced a short list of putative 174 male- and 258 female-specific P. falciparum stage V proteins. Furthermore, we generated antibodies against three putative female-specific gametocyte stage V proteins in P. falciparum and confirmed sex-specificity for two proteins and also the loss sex-partitioning for a putative female-specific protein in rodent malaria parasites.

Dataset Information

The translatome of quiescent Plasmodium falciparum gametocytes reveals parasite pyridoxal 5’-phosphate (PLP) biosynthesis is essential for efficient mosquito stage development

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets