Department of Applied Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi, 464-8601, Japan.
Department of Life Sciences, Graduate School of Bioresources, Mie University, 1577 Kurimamachiya‑cho, Tsu, Mie, 514‑8507, Japan.
Curr Genet. 2019 Aug;65(4):941-952. doi: 10.1007/s00294-019-00944-4. Epub 2019 Feb 22.
Cellulase production in filamentous fungi is repressed by various carbon sources. In our preliminary survey in Aspergillus nidulans, degree of de-repression differed depending on carbon sources in a mutant of creA, encoding the transcriptional repressor for carbon catabolite repression (CCR). To further understand mechanisms of CCR of cellulase production, we compared the effects of creA deletion with deletion of protein kinase A (pkaA) and G (ganB) genes, which constitute a nutrient sensing and signaling pathway. In plate culture with carboxymethyl cellulose and D-glucose, deletion of pkaA and ganB, but not creA, led to significant de-repression of cellulase production. In submerged culture with cellobiose and D-glucose or 2-deoxyglucose, both creA or pkaA single deletion led to partial de-repression of cellulase genes with the highest level by their double deletion, while ganB deletion caused de-repression comparable to that of the creA/pkaA double deletion. With ball-milled cellulose and D-glucose, partial de-repression was detected by deletion of creA but not of pkaA or ganB. The creA/pkaA or creA/ganB double deletion led to earlier expression than the creA deletion. Furthermore, the effect of each deletion with D-xylose or L-arabinose as the repressing carbon source was significantly different from that with D-glucose, D-fructose, and D-mannose. Consequently, this study revealed that PkaA and GanB participate in CreA-independent CCR and that contribution of CreA, PkaA, and GanB in CCR differs depending on the inducers, repressing carbon sources, and culture conditions (plate or submerged). Further study of CreA-independent mechanisms is needed to fully understand CCR in filamentous fungi.
纤维素酶在丝状真菌中的产生受到各种碳源的抑制。在我们对构巢曲霉中 creA 突变体的初步调查中,编码碳分解代谢物阻遏(CCR)转录阻遏物的 creA 基因的表达抑制程度因碳源而异。为了进一步了解纤维素酶产生的 CCR 机制,我们比较了 creA 缺失与蛋白激酶 A(pkaA)和 G(ganB)基因缺失的影响,这两个基因构成了营养感应和信号通路。在含有羧甲基纤维素和 D-葡萄糖的平板培养中,pkaA 和 ganB 的缺失,但不是 creA 的缺失,导致纤维素酶产生的显著去阻遏。在含有纤维二糖和 D-葡萄糖或 2-脱氧葡萄糖的液体培养中,creA 或 pkaA 的单一缺失导致纤维素酶基因的部分去阻遏,最高水平通过它们的双缺失,而 ganB 的缺失导致去阻遏与 creA/pkaA 双缺失相当。用球磨纤维素和 D-葡萄糖,通过 creA 的缺失检测到部分去阻遏,但 pkaA 或 ganB 的缺失则没有。creA/pkaA 或 creA/ganB 双缺失比 creA 缺失导致更早的表达。此外,用 D-木糖或 L-阿拉伯糖作为抑制碳源的每个缺失的影响与用 D-葡萄糖、D-果糖和 D-甘露糖的影响显著不同。因此,本研究表明 PkaA 和 GanB 参与 CreA 独立的 CCR,并且 CreA、PkaA 和 GanB 在 CCR 中的作用因诱导物、抑制碳源和培养条件(平板或液体)而异。需要进一步研究 CreA 独立的机制,以充分了解丝状真菌中的 CCR。