Figure 7
From: Metabolomics profiling reveals new aspects of dolichol biosynthesis in Plasmodium falciparum
Characterization of PfPPRD conditional knockdown mutant. (a) Schematic representation of the PfPPRD conditional knockdown mechanism. The modified PfPPRD locus contains a 3′-UTR RNA aptamer sequence that binds to TetR and DOZI fusion protein. In the presence of anhydrotetracycline (aTc), the TetR-DOZI complex is stabilized and the 3′-UTR aptamer is unbound. Removal of aTc causes binding of the TetR + DOZI repressor to the aptamer region which results in reduced protein levels. (b) Time-course of growth of PfPPRD-TetR-DOZI knockdown parasites treated with BSD only (− aTc) or BSD + aTc. Data are shown as mean ± SD (two independent experiments performed in triplicate) with error bars smaller than the dots. Individual values are shown in Supplementary Table S2. (*) Indicates p < 0.0001. A representative Giemsa-stained thin smear of parasites cultured in the presence or absence of aTc showing morphological defects observed in parasites after 8 days of aTc removal. The distribution of polyprenol (POH) and dolichol (DOH) species present in P. falciparum PfPPRD-TetR-DOZI were analyzed by LC-HRMS in trophozoite stage after aTc was removed from cultures for (c) 3 days and (d) 10 days. Values are average ± s.e.m. from three biological replicates. The figure was generated using CorelDraw 2018 (https://www.coreldraw.com/en/) and Adobe Photoshop 21.2.0 (https://www.adobe.com/products/photoshop.html).
