Wang Niyan, Yoshida Yusuke, Hasunuma Kohji
Kihara Institute for Biological Research, Yokohama City University, 641-12 Maioka-cho, Totsuka-ku, Yokohama, 244-0813, Japan.
Mol Genet Genomics. 2007 Jan;277(1):13-22. doi: 10.1007/s00438-006-0170-4. Epub 2006 Nov 1.
Light is one of the most important factors inducing morphogenesis in Neurospora crassa. The reception of light triggers the generation of reactive oxygen species (ROS) including hydrogen peroxide (H(2)O(2)). Catalase-1 (Cat-1) is one of three catalases known to detoxify H(2)O(2) into water and oxygen. We reported that the photomorphogenetic characteristics of mutants in nucleoside diphosphate kinase-1 (NDK-1), a light signal transducer, are severely affected, and NDK-1 interacted with Cat-1 in a yeast two-hybrid assay. To disclose the function of Cat-1, we created a Cat-1 loss-of-function mutant (cat-1 ( RIP )) by the repeat induced point-mutation (RIPing) method. No Cat-1 activity was detected in the mutant strain. Forty guanines were replaced with adenines in the cat-1 gene of cat-1 ( RIP ), which caused 30 amino acid substitutions. The mutant strain grew normally, but its conidia and mycelia were more sensitive to H(2)O(2) than those of the wild type. The lack of Cat-1 activity also caused a significant reduction in the conidial germination rate. Furthermore, light enhanced this reduction in cat-1 ( RIP ) more than that in the wild type. Introduction of cat-1 into the mutant reversed all of these defective phenotypes. These results indicate that Cat-1 plays an important role in supporting the survival of conidia under oxidative and light-induced stress.
光是以粗糙脉孢菌诱导形态发生的最重要因素之一。光的接收触发了包括过氧化氢(H₂O₂)在内的活性氧(ROS)的产生。过氧化氢酶-1(Cat-1)是已知的三种将H₂O₂解毒为水和氧气的过氧化氢酶之一。我们报道,作为光信号转导器的核苷二磷酸激酶-1(NDK-1)突变体的光形态发生特征受到严重影响,并且在酵母双杂交试验中NDK-1与Cat-1相互作用。为了揭示Cat-1的功能,我们通过重复诱导点突变(RIPing)方法创建了一个Cat-1功能丧失突变体(cat-1(RIP))。在突变菌株中未检测到Cat-1活性。在cat-1(RIP)的cat-1基因中,40个鸟嘌呤被腺嘌呤取代,这导致了30个氨基酸的替换。突变菌株生长正常,但其分生孢子和菌丝体比野生型对H₂O₂更敏感。Cat-1活性的缺乏也导致分生孢子萌发率显著降低。此外,光增强了cat-1(RIP)中这种降低的程度,比野生型更明显。将cat-1导入突变体可逆转所有这些缺陷表型。这些结果表明,Cat-1在支持分生孢子在氧化和光诱导应激下的存活中起重要作用。