Kawasaki L, Wysong D, Diamond R, Aguirre J
Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, D.F.
J Bacteriol. 1997 May;179(10):3284-92. doi: 10.1128/jb.179.10.3284-3292.1997.
Catalases are ubiquitous hydrogen peroxide-detoxifying enzymes that are central to the cellular antioxidant response. Of two catalase activities detected in the fungus Aspergillus nidulans, the catA gene encodes the spore-specific catalase A (CatA). Here we characterize a second catalase gene, identified after probing a genomic library with catA, and demonstrate that it encodes catalase B. This gene, designated catB, predicts a 721-amino-acid polypeptide (CatB) showing 78% identity to an Aspergillus fumigatus catalase and 61% identity to Aspergillus niger CatR. Notably, similar levels of identity are found when comparing CatB to Escherichia coli catalase HPII (43%), A. nidulans CatA (40%), and the predicted peptide of a presumed catA homolog from A. fumigatus (38%). In contrast, the last two peptides share a 79% identity. The catalase B activity was barely detectable in asexual spores (conidia), disappeared after germination, and started to accumulate 10 h after spore inoculation, throughout growth and conidiation. The catB mRNA was absent from conidia, and its accumulation correlated with catalase activity, suggesting that catB expression is regulated at the transcription level. In contrast, the high CatA activity found in spores was lost gradually during germination and growth. In addition to its developmental regulation, CatB was induced by H2O2, heat shock, paraquat, or uric acid catabolism but not by osmotic stress. This pattern of regulation and the protective role against H2O2 offered by CatA and CatB, at different stages of the A. nidulans life cycle, suggest that catalase gene redundancy performs the function of satisfying catalase demand at the two different stages of metabolic and genetic regulation represented by growing hyphae versus spores. Alternative H2O2 detoxification pathways in A. nidulans were indicated by the fact that catA/catB double mutants were able to grow in substrates whose catabolism generates H2O2.
过氧化氢酶是普遍存在的过氧化氢解毒酶,是细胞抗氧化反应的核心。在构巢曲霉中检测到的两种过氧化氢酶活性中,catA基因编码孢子特异性过氧化氢酶A(CatA)。在此,我们鉴定了第二个过氧化氢酶基因,该基因是在用catA探测基因组文库后确定的,并证明它编码过氧化氢酶B。这个基因命名为catB,预测编码一个721个氨基酸的多肽(CatB),与烟曲霉过氧化氢酶的同一性为78%,与黑曲霉CatR的同一性为61%。值得注意的是,将CatB与大肠杆菌过氧化氢酶HPII(43%)、构巢曲霉CatA(40%)以及烟曲霉假定的catA同源物的预测肽(38%)进行比较时,发现了相似的同一性水平。相比之下,后两种肽的同一性为79%。过氧化氢酶B的活性在无性孢子(分生孢子)中几乎检测不到,在萌发后消失,并在孢子接种10小时后开始积累,贯穿整个生长和产孢过程。分生孢子中不存在catB mRNA,其积累与过氧化氢酶活性相关,表明catB的表达在转录水平受到调控。相比之下,孢子中发现的高CatA活性在萌发和生长过程中逐渐丧失。除了其发育调控外,CatB还受到过氧化氢、热休克、百草枯或尿酸分解代谢的诱导,但不受渗透胁迫的诱导。这种调控模式以及CatA和CatB在构巢曲霉生命周期的不同阶段对过氧化氢的保护作用表明,过氧化氢酶基因冗余在以生长菌丝与孢子为代表的代谢和遗传调控的两个不同阶段发挥着满足过氧化氢酶需求的功能。构巢曲霉中存在替代的过氧化氢解毒途径,这一事实表明catA/catB双突变体能够在其分解代谢产生过氧化氢的底物中生长。