Wang Dan, Li Yingying, Wang Haichuan, Wei Dongsheng, Akhberdi Oren, Liu Yanjie, Xiang Biyun, Hao Xiaoran, Zhu Xudong
National Key Program of Microbiology and Department of Microbiology, College of Life Sciences, Nankai University (DMNU), Tianjin 300071, China.
Beijing Key Laboratory of Genetic Engineering Drug and Biotechnology, Institute of Biochemistry and Molecular Biology, College of Life Sciences, Beijing Normal University, Beijing 100875, China.
Genes (Basel). 2018 Jan 24;9(2):59. doi: 10.3390/genes9020059.
Highly conserved, the Snf1/AMPK is a central regulator of carbon metabolism and energy production in the eukaryotes. However, its function in filamentous fungi has not been well established. In this study, we reported functional characterization of Snf1/AMPK in the growth, development and secondary metabolism in the filamentous fungus . By deletion of the yeast homolog, we found that it regulated the utilization of carbon sources, e.g., sucrose, demonstrating a conserved function of this kinase in filamentous fungus. Importantly, several novel functions of were unraveled. For instance, the deletion strain displayed remarkable retardation in vegetative growth and pigmentation and produced a diminished number of conidia, even in the presence of the primary carbon source glucose. Deletion of the gene caused damages in the cell wall as shown by its hypersensitivities to Calcofluor white and Congo red, suggesting a critical role of Snf1 in maintaining cell wall integrity. Furthermore, the mutant strain Δ was hypersensitive to stress, e.g., osmotic pressure (1 M sorbitol), drug G418 and heat shock, though the mechanism remains to be illustrated. Significantly, disruption of the gene altered the production of secondary metabolites. By high-performance liquid chromatography (HPLC) profiling, we found that Δ barely produced secondary metabolites, e.g., the known product pestalotiollide B. This study suggests that Snf1 is a key regulator in filamentous fungus concerting carbon metabolism and the filamentous growth, conidiation, cell wall integrity, stress tolerance and the biosynthesis of secondary metabolites.
Snf1/AMPK高度保守,是真核生物中碳代谢和能量产生的核心调节因子。然而,其在丝状真菌中的功能尚未完全明确。在本研究中,我们报道了Snf1/AMPK在丝状真菌生长、发育和次级代谢中的功能特征。通过缺失其酵母同源物,我们发现它调节碳源(如蔗糖)的利用,证明了该激酶在丝状真菌中的保守功能。重要的是,还揭示了它的几个新功能。例如,缺失菌株在营养生长和色素沉着方面表现出显著延迟,分生孢子数量减少,即使在存在主要碳源葡萄糖的情况下也是如此。该基因的缺失导致细胞壁受损,表现为对荧光增白剂和刚果红高度敏感,表明Snf1在维持细胞壁完整性方面起关键作用。此外,突变菌株Δ对压力(如渗透压(1 M山梨醇)、药物G418和热休克)高度敏感,尽管其机制尚待阐明。值得注意的是,该基因的破坏改变了次级代谢产物的产生。通过高效液相色谱(HPLC)分析,我们发现Δ几乎不产生次级代谢产物,如已知产物pestalotiollide B。本研究表明,Snf1是丝状真菌中协调碳代谢与丝状生长、分生孢子形成、细胞壁完整性、胁迫耐受性和次级代谢产物生物合成的关键调节因子。