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The P. falciparum orthologue of Male Development Protein 3 is a Male-associated regulator of translation initiation or enhancement [CLIP-Seq]

ABSTRACT: Sex determination and development of Plasmodium gametocytes is critical to the transmission of malaria parasites and involves a complex interplay of multiple molecular factors. Here, we characterized the P. falciparum gene syntenic to P. berghei MD3- pf3d7_0315600. This protein contains a non-classical CCCH zinc-finger domain and Lysine-Proline rich domain associated with ribosome receptors, suggesting a role in mRNA regulation. We investigated the function of PfMD3-3xHA using in vivo using crosslinked RNA-Protein interaction capture followed by RNA-sequencing (CLIP-seq) to determine its RNA targets in both early (stage II) and late (stage IV-V) development of gametocytes. We found that these bound mRNA targets show decreased protein levels in a genetic knockout of PfMD3, suggesting a role for PfMD3 in translational processing. We then propose that PfMD3 functions as a Male Associated Regulator of Translation Initiation or Enhancement (MARTIE) in the human malaria parasite P. falciparum.

ORGANISM(S): Plasmodium falciparum

PROVIDER: GSE267362 | GEO | 2024/09/01

REPOSITORIES: GEO

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Project description:For malaria transmission, the parasite must undergo sexual differentiation into mature gametocytes. However, the molecular basis for this critical transition in the parasites life cycle is unknown. Six previously uncharacterized genes, Pfg14.744, Pfg14.745, Pfg14.748, Pfg14.763, Pfg14.752 and Pfg6.6 that are members of a 36 gene Plasmodium falciparum-specific subtelomeric superfamily were found to be expressed in parasites that are committed to sexual development as suggested by co-expression of Pfs16 and Pfg27. Northern blots demonstrated that Pfg14.744 and Pfg14.748 were first expressed before the parasites differentiated into morphologically distinct gametocytes, transcription continued to increase until stage II gametocytes were formed and then rapidly decreased. Immunofluorescence assays indicated that both proteins were only produced in the subpopulation of ring stage parasites that are committed to gametocytogenesis and both localized to the parasitophorous vacuole (PV)b of the early ring stage parasites. As the parasites continued to develop Pfg14.748 remained within the parasitophorous vacuole, while Pfg14.744 was detected in the erythrocyte. The 5' flanking region of either gene alone was sufficient to drive early gametocyte specific expression of green fluorescent protein (GFP). In parasites transfected with a plasmid containing the Pfg14.748 5' flanking region immediately upstream of GFP, fluorescence was observed in a small number of schizonts the cycle before stage I gametocytes were observed. This expression pattern is consistent with commitment to sexual differentiation prior to merozoite release and erythrocyte invasion. Further investigation into the role of these genes in the transition from asexual to sexual differentiation could provide new strategies to block malaria transmission. Microarray analysis was used to compare two clones derived from Plasmodium falciparum strain 3D7 parasites that differ in their ability to undergo gametocytogenesis. Clone G+ produces gametocytes and clone G- produces very few if any gametocytes. RNA was harvested from the cultures when the asexual parasitemia was 0.9-1.48% (day 4) (n=4) after setting up the gametocyte cultures and 5.2-5.58% (day 6) (n=4) prior to the appearance of morphologically distinct gametocytes and used to generate cDNA that was labeled with Cy3 or Cy5 and hybridized to the Plasmodium falciparum 70 mer oligonucleotide microarray developed by DeRisi and co-workers.

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The P. falciparum orthologue of Male Development Protein 3 is a Male-associated regulator of translation initiation or enhancement [CLIP-Seq]

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