Dataset Information

Mapping PP1c and its Inhibitor 2 interactomes reveals conserved and specific networks in asexual and sexual stages of Plasmodium

ABSTRACT: Malaria parasites require multiple phosphorylation and dephosphorylation steps to drive signaling pathways for proper differentiation and transformation. Several protein phosphatases, including Protein Phosphatase 1 (PP1c), one of the main dephosphorylation enzymes, have been shown to be indispensable for Plasmodium life cycle. PP1c functions are elaborated processes via dynamic interactions with a vast number of binding partners that contribute to its diversity of action. In this study, we have used Plasmodium berghei transgenic parasite strains stably expressing PP1c or its Inhibitor 2 (I2) tagged with mCherry, combined with mCherry affinity pulldown of proteins from asexual and sexual stages and mass spectrometry analyses. Mapped proteins were used to identify interactomes and to cluster functionally related proteins. Our findings confirm previous physical interactions of PP1c and reveal shared and specific biological processes enrichment linked to cellular component assembly in both schizonts and gametocytes, to biosynthetic process/translation in schizonts and to protein transport exclusively in gametocytes. Further, our analysis of PP1c and I2 interactomes reveals that nuclear export mediator factor and peptidyl-prolyl cis-trans isomerase, suggested to be essential in P. falciparum, could be potential targets of the complex PP1c/I2 in both asexual and sexual stages. Our study emphasizes the adaptability of Plasmodium PP1c and provides a fundamental study of the protein interaction landscapes involved in a myriad of events in Plasmodium, suggesting why it is crucial to the parasite and a source for alternative therapeutic strategies.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Plasmodium Berghei Anka Mus Musculus (mouse)

TISSUE(S): Blood

SUBMITTER: Cerina Chhuon

LAB HEAD: Chiara Guerrera

PROVIDER: PXD029283 | Pride | 2022-02-22

REPOSITORIES: Pride

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Publications

Mapping PP1c and Its Inhibitor 2 Interactomes Reveals Conserved and Specific Networks in Asexual and Sexual Stages of <i>Plasmodium</i>.

De Witte Caroline C Aliouat El Moukhtar EM Chhuon Cerina C Guerrera Ida Chiara IC Pierrot Christine C Khalife Jamal J

International journal of molecular sciences 20220119 3

Malaria parasites require multiple phosphorylation and dephosphorylation steps to drive signaling pathways for proper differentiation and transformation. Several protein phosphatases, including protein phosphatase 1 (PP1), one of the main dephosphorylation enzymes, have been shown to be indispensable for the <i>Plasmodium</i> life cycle. The catalytic subunit of PP1 (PP1c) participates in cellular processes via dynamic interactions with a vast number of binding partners that contribute to its di ...[more]

PMID: 35162991

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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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Dataset Information

Mapping PP1c and its Inhibitor 2 interactomes reveals conserved and specific networks in asexual and sexual stages of Plasmodium

Publications

Mapping PP1c and Its Inhibitor 2 Interactomes Reveals Conserved and Specific Networks in Asexual and Sexual Stages of <i>Plasmodium</i>.

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