2026-10-12T00:53:23Z https://api.figshare.com/v2/oai

oai:figshare.com:article/34017863 2026-09-28T17:55:03Z category_1 category_4 category_915 category_7 category_8 category_12 category_13 category_132 portal_5 item_type_1 month_year_09_2026

PfATG18 is critical for V-ATPase integrity. Yasmin Schmitz (11974910) Moumita Sengupta (25132859) Carola Schneider (3743791) Tanja Ziesmann (3318936) Franziska Hellmold (25132862) Ute Distler (115774) Rudolph Reimer (190562) Joachim Michael Matz (25132865) Biophysics Biochemistry Physical Sciences not elsewhere classified Medicine Microbiology Cell Biology Genetics Infectious Diseases related protein 18 div >< p broad architectural destabilization balance cargo influx associated protein machinery antimalarial drugs unchanged 3d electron microscopy membrane sector phenocopied endocytic membrane homeostasis endocytic membrane dynamics vacuole constantly fuses specialized digestive vacuole stage parasites hypersensitive vacuolar membrane dynamics efficient hemoglobin processing degrading hemoglobin within asexual parasite proliferation plasmodium falciparum </ atpase assembly factor atg18 links v vacuolar membrane vacuole ’ malaria parasites atpase assembly parasite ’ parasite survival parasite death incoming hemoglobin pf </ vesicle trafficking rendered ring regulatory influence quantitative live proton pump point mutation multifaceted role key regulator interactions indicates interact directly findings establish filled vesicles direct interference decrease sensitivity (A) Schematic of the V-ATPase complex, with sectors and relevant subunits indicated. (B,C) Subunit V1B dissociates from the DVM upon PfATG18 loss. (B) Live colocalization of CRT-mCh and V1B-mNG. Yellow arrows: transects for intensity plots; light gray: parasite cytoplasm; dark gray: DV; black arrowheads: DVM. (C) Ratio of DVM- to cytosol-localized V1B-mNG intensity. Individual and mean values; paired t-test; n = 99 parasites, 3 experiments. *, p < 0.05. Parasites were induced at ring stage and imaged at 36 hours post-invasion (hpi). The dashed line denotes full equilibration of both V1B pools. (D,E) Subunit Voa is strongly depleted from the DVM in PfATG18-deficient parasites. Analysis as in (B,C). (F) V1B dissociation precedes DV fragmentation. Loss of PfATG18 was induced at 28 hpi, shifting the onset of DV fragmentation to 36 hpi. Analysis as in (C). Individual and mean values; one-way ANOVA with Tukey’s test; n = 165 parasites, 5 experiments. n.s., p > 0.05; **, p < 0.01. (G) Conditional inactivation of subunit Voc recapitulates the PfATG18 knockout phenotype, with DV fission and HTC accumulation (arrowheads). Shown are CRT-mNG and the first 180 amino acids of glycophorin-binding protein 130 fused to mSc (GBP1301-180-mSc), an exported and reinternalized marker. (H,I) Co-immunoprecipitation combined with mass spectrometry identifies candidate PfATG18 interaction partners involved in phosphoinositide balance and membrane trafficking. (H) Protein hits ranked by enrichment score, defined as the log2 fold-change (atg18-mNG:loxP/B11) multiplied by the negative log10 of the p-value. Benjamini-Hochberg corrected t-test; n = 3 experiments with 3 technical replicates each. Relevant proteins as well as unrelated DVM markers are indicated. (I) Membrane trafficking-related proteins among the top 200 hits with a confirmed or potential role in the endolysosomal pathway. Scale bars: 5 µm. Source data are provided as a <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1014608#ppat.1014608.s015" target="_blank">S3 File</a>. 2026-09-28T18:08:16Z Image Figure 10.1371/journal.ppat.1014608.g006 https://figshare.com/articles/figure/_p_i_Pf_i_ATG18_is_critical_for_V-ATPase_integrity_p_/34017863 CC BY 4.0