Molina Lázaro, Kahmann Regine
Max Planck Institute for Terrestrial Microbiology, D-35043 Marburg, Germany.
Plant Cell. 2007 Jul;19(7):2293-309. doi: 10.1105/tpc.107.052332. Epub 2007 Jul 6.
The fungus Ustilago maydis is a biotrophic pathogen of maize (Zea mays). In its genome we have identified an ortholog of YAP1 (for Yeast AP-1-like) from Saccharomyces cerevisae that regulates the oxidative stress response in this organism. yap1 mutants of U. maydis displayed higher sensitivity to H(2)O(2) than wild-type cells, and their virulence was significantly reduced. U. maydis yap1 could partially complement the H(2)O(2) sensitivity of a yap1 deletion mutant of S. cerevisiae, and a Yap1-green fluorescent protein fusion protein showed nuclear localization after H(2)O(2) treatment, suggesting that Yap1 in U. maydis functions as a redox sensor. Mutations in two Cys residues prevented accumulation in the nucleus, and the respective mutant strains showed the same virulence phenotype as Deltayap1 mutants. Diamino benzidine staining revealed an accumulation of H(2)O(2) around yap1 mutant hyphae, which was absent in the wild type. Inhibition of the plant NADPH oxidase prevented this accumulation and restored virulence. During the infection, Yap1 showed nuclear localization after penetration up to 2 to 3 d after infection. Through array analysis, a large set of Yap1-regulated genes were identified and these included two peroxidase genes. Deletion mutants of these genes were attenuated in virulence. These results suggest that U. maydis is using its Yap1-controlled H(2)O(2) detoxification system for coping with early plant defense responses.
玉米黑粉菌(Ustilago maydis)是玉米(Zea mays)的一种活体营养型病原体。在其基因组中,我们鉴定出了酿酒酵母(Saccharomyces cerevisae)中YAP1(酵母AP-1样蛋白)的一个直系同源物,该蛋白在酿酒酵母中调节氧化应激反应。玉米黑粉菌的yap1突变体对H₂O₂的敏感性高于野生型细胞,且其毒力显著降低。玉米黑粉菌的Yap1可以部分弥补酿酒酵母yap1缺失突变体对H₂O₂的敏感性,并且Yap1-绿色荧光蛋白融合蛋白在H₂O₂处理后显示出核定位,这表明玉米黑粉菌中的Yap1作为一种氧化还原传感器发挥作用。两个半胱氨酸残基的突变阻止了其在细胞核中的积累,相应的突变菌株表现出与Δyap1突变体相同的毒力表型。二氨基联苯胺染色显示,yap1突变体菌丝周围积累了H₂O₂,而野生型中不存在这种情况。抑制植物NADPH氧化酶可阻止这种积累并恢复毒力。在感染过程中,Yap1在感染后2至3天穿透后显示出核定位。通过阵列分析,鉴定出了大量受Yap1调控的基因,其中包括两个过氧化物酶基因。这些基因的缺失突变体毒力减弱。这些结果表明,玉米黑粉菌利用其由Yap1控制的H₂O₂解毒系统来应对植物早期的防御反应。