Michán Shaday, Lledías Fernando, Hansberg Wilhelm
Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, DF, México.
Eukaryot Cell. 2003 Aug;2(4):798-808. doi: 10.1128/EC.2.4.798-808.2003.
We use asexual development of Neurospora crassa as a model system with which to determine the causes of cell differentiation. Air exposure of a mycelial mat induces hyphal adhesion, and adherent hyphae grow aerial hyphae that, in turn, form conidia. Previous work indicated the development of a hyperoxidant state at the start of these morphogenetic transitions and a large increase in catalase activity during conidiation. Catalase 3 (CAT-3) increases at the end of exponential growth and is induced by different stress conditions. Here we analyzed the effects of cat-3-null strains on growth and asexual development. The lack of CAT-3 was not compensated by other catalases, even under oxidative stress conditions, and cat-3(RIP) colonies were sensitive to H(2)O(2), indicating that wild-type (Wt) resistance to external H(2)O(2) was due to CAT-3. cat-3(RIP) colonies grown in the dark produced high levels of carotenes as a consequence of oxidative stress. Light exacerbated oxidative stress and further increased carotene synthesis. In the cat-3(RIP) mutant strain, increased aeration in liquid cultures led to increased hyphal adhesion and protein oxidation. Compared to the Wt, the cat-3(RIP) mutant strain produced six times more aerial hyphae and conidia in air-exposed mycelial mats, as a result of longer and more densely packed aerial hyphae. Protein oxidation in colonies was threefold higher and showed more aerial hyphae and conidia in mutant strains than did the Wt. Results indicate that oxidative stress due to lack of CAT-3 induces carotene synthesis, hyphal adhesion, and more aerial hyphae and conidia.
我们利用粗糙脉孢菌的无性发育作为一个模型系统来确定细胞分化的原因。将菌丝体垫暴露于空气中会诱导菌丝黏附,而黏附的菌丝会长出气生菌丝,气生菌丝进而形成分生孢子。先前的研究表明,在这些形态发生转变开始时会出现高氧化态,并且在分生孢子形成过程中过氧化氢酶活性大幅增加。过氧化氢酶3(CAT-3)在指数生长末期增加,并由不同的应激条件诱导产生。在此,我们分析了cat-3基因缺失菌株对生长和无性发育的影响。即使在氧化应激条件下,其他过氧化氢酶也无法补偿CAT-3的缺失,并且cat-3(RIP)菌落对H₂O₂敏感,这表明野生型(Wt)对外部H₂O₂的抗性归因于CAT-3。在黑暗中生长的cat-3(RIP)菌落由于氧化应激产生了高水平的类胡萝卜素。光照加剧了氧化应激并进一步增加了类胡萝卜素的合成。在cat-3(RIP)突变菌株中,液体培养中通气增加导致菌丝黏附增加和蛋白质氧化。与野生型相比,cat-3(RIP)突变菌株在暴露于空气的菌丝体垫中产生的气生菌丝和分生孢子是野生型的六倍,这是由于气生菌丝更长且更密集。突变菌株菌落中的蛋白质氧化比野生型高两倍,并且显示出更多的气生菌丝和分生孢子。结果表明,由于缺乏CAT-3导致的氧化应激会诱导类胡萝卜素合成、菌丝黏附以及更多的气生菌丝和分生孢子。