Secondary structure of Pf mt rRNA fragments and their likely positions in the modeled SSU and LSU.

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PfRSM22 and PfMRPL23 are localized to the mitochondrion and are essential for parasite survival.

(A) Images from fluorescence microscopy showing colocalization of PfRSM22-3HA (green) with Mitotracker (red). (B) Colocalization of PfMRPL23-3HA (green) with Mitotracker (red). In A and B, the parasite nucleus is stained using DAPI (blue). (C) Western blot showing expression of PfRSM22-3HA protein (62 KDa) in the presence or absence of 250 nM aTc over 6 days (3 IDCs). (D) Western blot showing expression of PfMRPL23-3HA protein (32 KDa) in the presence or absence of 250 nM aTc over 4 days (2 IDCs). Pf-Exp2 (33 KDa) was used as a loading control. Representative images of Giemsa-stained D10-PfRSM22-3HA parasites (E) and D10-PfMRPL23-3HA parasites (F) at the trophozoite stage in the presence (aTc ON) and absence of aTc (aTc OFF). Quantification of D10-PfRSM22-3HA parasite growth (G) and D10-PfMRPL23-3HA parasite growth (H) over 8 days in the presence and absence of aTc. Growth index was calculated by multiplication of parasitemia with splitting factors over the time course. Data shown here is mean ± SD of n = 3.</p

Late effects of PfRSM22 and PfMRPL23 KDs on transcripts of mitoribosomal components.

(A) Log2 fold change of putative PfMRPs on day 6 after PfRSM22 KD (orange) and on day 4 after PfMRPL23 KD (blue). (B) Log2 fold change of mt rRNA fragments on day 6 after PfRSM22 KD (orange) and on day 4 after PfMRPL23 KD (blue).</p

Upregulated non-mitochondrial transcripts common between PfRSM22 and PfMRPL23 KDs in the late phase.

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PfRSM22 and PfMRPL23 KD increased sensitivity to antimalarials targeting the parasite <i>bc1</i> complex.

(A) The schematic represents key steps of growth inhibition assays measured by [3H] hypoxanthine incorporation. The diagram was created using BioRender.com. (B and C) Atovaquone hypersensitivity upon PfRSM22 and PfMRPL23 KD respectively. (D and E) ELQ-300 hypersensitivity upon PfRSM22 and PfMRPL23 KD respectively. Reduction in IC50 values and Hill Slope of the curve were reported for respective parasite lines. Data shown are the mean ± S.D. of triplicates from n = 3 independent experiments. (F and G) KD of PfRSM22 or PfMRPL23 did not show hypersensitivity to chloroquine.</p

RT-qPCR of representative SSU and LSU mt rRNA upon protein KD.

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S1 File -

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PfMRPL23 protein domain and amino acid sequence alignment.

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Early and late effects of PfRSM22 and PfMRPL23 KD on apicoplast related transcripts.

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Downregulated non-mitochondrial transcripts common between PfRSM22 and PfMRPL23 KDs in the late phase.

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