Fig. 6: Molecular interactions and functional roles of Ntp1 and its truncated variants in aflatoxin biosynthesis and developmental processes in A. flavus.
From: The role of Npt1 in regulating antifungal protein activity in filamentous fungi
A, B Analysis of the interaction between AfAFP and PgAFP with various Ntp1 mutant proteins. Top: Western blot detection with anti-HA antibodies in whole-cell lysates of A. flavus expressing HA-tagged Ntp1 mutant proteins. Middle: Western blots of immunoprecipitates with anti-HA antibody from the corresponding lysates. Bottom: Strep-Tactin beads loaded with purified AfAFP-F-S, PgAFP-F-S, and F-S proteins. C In silico docking simulations using AlphaFold2 multimer to analyze interactions between Ntp1546–588 and PgAFP with AfAFP. The simulations revealed that for Ntp1546–588 and PgAFP, there were three hydrogen bonds (two between Ntp1V579 and PgAFPV25, and one between Ntp1F581 and PgAFPN23) and one salt bridge (between Ntp1K588 and PgAFPE7). For Ntp1546–588 and AfAFP, there were four hydrogen bonds (two between Ntp1V579 and AfAFPF26, one between Ntp1F581 and AfAFPD24, and another between Ntp1N582 and AfAFPK23). The visual representations of PgAFP and AfAFP are depicted as cartoons in yellowish brown and light red, respectively, while the structure of Ntp1546–588 is shown in light blue. The predicted protein-protein interaction structures are also categorized by four levels of confidence intervals. D Growth phenotypes of WT, Δntp1285–708, Δntp1417–708, and Δntp1589–708 strains on YGT medium. E Colony diameter measurements of the strains. F Microscopic view of conidiophore development in the WT, Δntp1285–708, Δntp1417–708, and Δntp1589–708 strains. G Spore counts of the strains. H Sclerotia formation phenotypes of WT, Δntp1285–708, Δntp1417–708, and Δntp1589–708 strains on CM medium. I The amount of sclerotia generated by these strains. J Aflatoxin production in WT, Δntp1285–708, Δntp1417–708, and Δntp1589–708 mutant strains, as detected by thin-layer chromatography (TLC). K Optical density-based semi-quantification of aflatoxin levels corresponding to (J). Data in (E, G, I, K) are presented as mean ± SEM (n ≥ 3). Statistical significance was determined by one-way ANOVA and indicated by *** for P < 0.001. All experiments were performed in triplicate. Source data are provided as a Source Data file.
