Figure 3
FvC5SD is an iron regulated gene involved in iron uptake.
(a) Spot dilution assay of WT strain of S. cerevisiae (SC4741), erg3 knockout strain with FvC5SD cloned in pGal10HO vector or empty vector on synthetic minimal medium without and with 100μM BPA (limiting iron medium). (b) Northern blot analysis to show iron mediated regulation of FvC5SD transcription. Total RNA was isolated from F. velutipes mycelia grown in medium with or without BPA (100 μM). Graph shows mRNA levels on autoradiograms quantified by densitometry scanning using Fluor-S-(Bio-Rad) software. Data are mean ± SE (n = 3). (c) Comparison of radioactive iron (Fe55Cl3) uptake by S. pombe strain expressing FvC5SD and empty pSL173 vector at different time points. Data are mean ± SE (n = 3). (d) FvC5sdp structure predicted by homology based modelling using ESyPred3D server and threading by LOOP server. The two probable iron binding sites for each structure are also shown. Arrow is used to depict the residue in the iron binding sites which is also part of histidine rich motifs. WT, wild type; BPA, Bathophenanthrolinedisulphonic acid (iron chelator).
