Nakamura Hidetoshi, Kikuma Takashi, Jin Feng Jie, Maruyama Jun-ichi, Kitamoto Katsuhiko
Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
J Biosci Bioeng. 2016 Apr;121(4):365-71. doi: 10.1016/j.jbiosc.2015.08.011. Epub 2015 Oct 20.
The serine-threonine kinase Rim15p is a master regulator of stress signaling and is required for stress tolerance and sexual sporulation in the yeast Saccharomyces cerevisiae. However, in filamentous fungi that reproduce asexually via conidiation, the physiological function of Rim15p homologs has not been extensively analyzed. Here, we functionally characterized the protein homolog of Rim15p in the filamentous fungus Aspergillus oryzae, by deleting and overexpressing the corresponding Aorim15 gene and examining the role of this protein in stress tolerance and development. Deletion of Aorim15 resulted in an increase in the sensitivity of conidia to oxidative and heat stresses, whereas conidia of the Aorim15 overexpressing strain were more resistant to these stresses. These results indicated that AoRim15 functions in stress tolerance, similar to S. cerevisiae Rim15p. Phenotypic analysis revealed that conidiation was markedly reduced by overexpression of Aorim15 in A. oryzae, and was completely abolished in the deletion strain. In addition, the formation of sclerotia, which is another type of developmental structure in filamentous fungi, was decreased by the deletion of Aorim15, whereas Aorim15 overexpression increased the number of sclerotia. These results indicated that AoRim15 is a positive regulator of sclerotia formation and that overexpression of AoRim15 shifts the developmental balance from conidiation towards sclerotia formation. Collectively, we demonstrated that AoRim15 is involved in the stress tolerance of conidia and differentially regulates between the two developmental fates of conidiation and sclerotia formation.
丝氨酸 - 苏氨酸激酶Rim15p是应激信号的主要调节因子,对于酿酒酵母(Saccharomyces cerevisiae)的应激耐受性和有性孢子形成是必需的。然而,在通过分生孢子进行无性繁殖的丝状真菌中,Rim15p同源物的生理功能尚未得到广泛分析。在这里,我们通过删除和过表达丝状真菌米曲霉(Aspergillus oryzae)中相应的Aorim15基因,并研究该蛋白在应激耐受性和发育中的作用,对Rim15p的蛋白同源物进行了功能表征。删除Aorim15导致分生孢子对氧化应激和热应激的敏感性增加,而过表达Aorim15菌株的分生孢子对这些应激更具抗性。这些结果表明,AoRim15在应激耐受性中发挥作用,类似于酿酒酵母的Rim15p。表型分析表明,米曲霉中Aorim15的过表达显著降低了分生孢子形成,而在缺失菌株中则完全消除。此外,丝状真菌中另一种发育结构菌核的形成因Aorim15的缺失而减少,而Aorim15的过表达增加了菌核的数量。这些结果表明,AoRim15是菌核形成的正调节因子,并且AoRim15的过表达将发育平衡从分生孢子形成转向菌核形成。总的来说,我们证明了AoRim15参与分生孢子的应激耐受性,并在分生孢子形成和菌核形成这两种发育命运之间进行差异调节。