Extended Data Fig. 4: NCR247 drives iron uptake by controlling Irr mediated iron regulation.
From: A haem-sequestering plant peptide promotes iron uptake in symbiotic bacteria
a, Fluorescence of FITC-NCR247 quenched by increasing concentrations of heme. Fluorescence of FITC-NSR247 remains unquenched even after addition of excess heme. b-f, Decrease in expression of genes involved in iron uptake (fhuP (b), fbpA (c), rhrA (d), hmuP (e), foxA (f)) in a 2 µM NCR247 treated Δirr when compared to NCR247 treated wild type S. meliloti, when grown in iron-replete medium (30 µM). NCR247 was treated for 30 mins g and h, Growth pattern of untreated(g) and NCR247 treated (h) wildtype and Δirr cells in iron-depleted (-Fe) and iron-replete media (30 µM). i, Derepressed expression of hmuP in an untreated ΔrirA when compared to wildtype S. meliloti as measured by qRT-PCR analysis. j, Increased uptake of 55Fe in untreated ΔrirA when compared to untreated and NCR247 treated wildtype S. meliloti. k and l, Growth pattern of untreated (k) and NCR247 treated (l) wildtype and ΔrirA cells in iron-depleted (-Fe), iron sufficient (5 µM) and iron-replete media (30 µM). In a, g, h, j, k and l data are presented as mean of three independent replicates ± s.d. In b-f and i, The data are expressed as starting quantities (SQ) of respective mRNAs normalized to the control gene SMc00128 and are presented as average of three technical replicates ± s.d. In b-f, ****P < 0.0001 NCR247 treated WT vs Δirr - samples; In i, ***P = 0.0003 WT untreated vs WT NCR247 treated; two-way analysis of variance (ANOVA) with multiple comparisons.
