5 times more mRNA accumulation of the rcnA gene when mycelia were exposed to 0.5 mM menadione compared to mycelia not exposed to it (data not shown). Figure 6 Molecular characterization of the A. nidulans AnrcnA gene. (A) Schematic illustration of the

AnrcnA deletion strategy. (A) Genomic DNA from both wild type and ΔAnrcnA strains was SHP099 solubility dmso isolated and cleaved with the enzyme EcoRI; a 2.0-kb DNA fragment from the 3′-noncoding region was used as a hybridization probe. This fragment recognizes a single DNA band (about 10.7-kb) click here in the wild type strain and also a single DNA band (about 5.2-kb) in the ΔAnrcnA mutant as shown in the Southern blot analysis. (B) Wild type and ΔAnrcnA mutant strains were grown for 72 hours at 37°C in complete medium in the absence or presence of cyclosporine A 250 ng/ml and paraquat Doramapimod concentration 4 mM. (C) Growth phenotypes of A. nidulans wild type, ΔAnrcnA, ΔAncnaA, ΔAncnaA mutant strains were grown in complete medium for 72 hours at 37°C. In (B) and (C) graphs show the radial growth (cm) of the strains under different growth conditions. The results are the means ± standard deviation of four sets of experiments. (D) GFP::AnRcnA localizes to the cytoplasm. Germlings of the

GFP::AnRcnA were grown in liquid MM+ 2% glycerol for 24 hs at 30°C. The germlings were treated or not with 50 mM calcium chloride for different periods of time from 5 to 60 minutes. After the treatment, germlings were analysed by laser scanning confocal microscopy. The figure shows a GFP::AnRcnA germling exposed to calcium chloride; however, germlings not exposed to calcium chloride displayed essentially the same results. Images were captured by direct acquisition. Bars, 5 μm. The first member identified from the calcipressin family, RCAN1, was isolated from the hamster genome as a gene induced during transient adaptation to oxidative stress [42, 43]. It was observed that resistance to oxidative stress and calcium stress increased as a function of RCAN1 expression and decreased as its expression diminished [44]. Porta et al. [35] have shown that RCAN1

mRNA and protein expression are sensitive to oxidative stress in primary neurons, and that Rcan1 -/- neurons display an increased resistance to damage by hydrogen peroxide. Taken together, our results suggest that Aspergilli RcnA play a role in calcium and Mannose-binding protein-associated serine protease oxidative stress signaling. Next step, we crossed the A. nidulans ΔAnrcnA strain with ΔAncnaA strain (cnaA encodes the catalytic subunit of the calcineurin gene) [30]. The A. nidulans ΔAnrcnA mutation can partially suppress the ΔAncnaA growth defect, suggesting a genetic interaction between AnRcnA and AnCnaA (Figure 6C). To determine the AnRcnA cellular localization, we transformed a GFP::AnRcnA cassette into a wild type strain. Several transformants were obtained in which the plasmid had integrated homologously at the AnrcnA locus (data not shown).