Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Centre for Microbiology; College of Life Sciences, Nanjing Normal Universitygrid.260474.3, Nanjing, China.
mBio. 2021 Dec 21;12(6):e0300721. doi: 10.1128/mBio.03007-21. Epub 2021 Nov 16.
The life cycle of filamentous fungi generally comprises hyphal growth and asexual reproduction. Both growth and propagation processes are critical for invasion growth, spore dissemination, and virulence in fungal pathogens and for the production of secondary metabolites or for biomass accumulation in industrial filamentous fungi. The CAAT-inding omplex (CBC) is a heterotrimeric transcription factor comprising three subunits, HapB, HapC, and HapE, and is highly conserved in fungi. Previous studies revealed that CBC regulates sterol metabolism by repressing several genes in the ergosterol biosynthetic pathway in the human fungal pathogen Aspergillus fumigatus. In the present study, we found dysfunction of CBC caused the abnormal asexual reproduction (conidiation) in submerged liquid culture. CBC suppresses the activation of the gene in the central regulatory pathway for conidiation combined with its upstream regulators , , and by binding to the 5'-CCAAT-3' motif within conidiation gene promoters, and lack of CBC member HapB results in the upregulation of these genes. Furthermore, when the expression of or is repressed, the submerged conidiation does not happen in the mutant. Interestingly, deletion of HapB leads to enhanced transient cytosolic Ca levels and activates conidiation-positive inducer Ca-CrzA modules to enhance submerged conidiation, demonstrating that CrzA works with CBC as a reverse regulator of fungal conidiation. To the best of our knowledge, the finding of this study is the first report for the molecular switch mechanism between vegetative hyphal growth and asexual development regulated by CBC, in concert with Ca-CrzA signaling in A. fumigatus. A precisely timed switch between vegetative hyphal growth and asexual development is a crucial process for the filamentous fungal long-term survival, dissemination, biomass production, and virulence. However, under the submerged culture condition, filamentous fungi would undergo constant vegetative growth whereas asexual conidiation rarely occurs. Knowledge about possible regulators is scarce, and how they could inhibit conidiation in liquid culture is poorly understood. Here, we demonstrated that the transcription factor heterotrimeric CBC dominantly maintains vegetative growth in liquid-submerged cultures by directly suppressing the conidiation-inductive signal. In contrast, calcium and the transcription factor CrzA, are positive inducers of conidiation. Our new insights into the CBC and Ca-CrzA regulatory system for transition control in the submerged conidiation of A. fumigatus may have broad repercussions for all filamentous fungi. Moreover, our elucidation of the molecular mechanism for submerged conidiation may support new strategies to precisely control vegetative growth and asexual conidiation in aspergilli used in industry.
丝状真菌的生命周期通常包括菌丝生长和无性繁殖。生长和繁殖过程对于真菌病原体的入侵生长、孢子传播和毒力以及工业丝状真菌中次生代谢物的产生或生物量积累都至关重要。CAAT 结合复合物(CBC)是一种异三聚体转录因子,由三个亚基 HapB、HapC 和 HapE 组成,在真菌中高度保守。先前的研究表明,CBC 通过抑制人类真菌病原体烟曲霉中甾醇生物合成途径中的几个基因来调节固醇代谢。在本研究中,我们发现 CBC 功能障碍导致其在液体浸没培养中无性繁殖(分生孢子形成)异常。CBC 通过结合到分生孢子基因启动子中的 5'-CCAAT-3' 基序,抑制中央调控途径中基因的激活,与上游调节剂 、 和 共同作用,缺乏 CBC 成员 HapB 导致这些基因的上调。此外,当 或 的表达受到抑制时, 突变体中不会发生液体浸没分生孢子形成。有趣的是,HapB 的缺失导致细胞质 Ca 水平短暂升高,并激活分生孢子阳性诱导剂 Ca-CrzA 模块,以增强液体浸没分生孢子形成,表明 CrzA 与 CBC 一起作为真菌分生孢子形成的反向调节剂。据我们所知,这项研究的发现是第一个报道 CBC 调节的营养菌丝生长和无性发育之间的分子开关机制的报告,与烟曲霉中的 Ca-CrzA 信号协同作用。营养菌丝生长和无性发育之间的精确时间切换是丝状真菌长期生存、传播、生物量产生和毒力的关键过程。然而,在液体浸没培养条件下,丝状真菌将经历持续的营养生长,而无性分生孢子形成很少发生。关于可能的调节剂的知识很少,并且它们如何在液体培养中抑制分生孢子形成的知之甚少。在这里,我们证明转录因子异三聚体 CBC 通过直接抑制分生孢子诱导信号,主要维持液体浸没培养中的营养生长。相比之下,钙和转录因子 CrzA 是分生孢子形成的正诱导剂。我们对 CBC 和 Ca-CrzA 调控系统在烟曲霉浸没分生孢子形成中的转换控制的新见解可能对所有丝状真菌都有广泛的影响。此外,我们对浸没分生孢子形成的分子机制的阐明可能为在工业中使用的曲霉中精确控制营养生长和无性分生孢子形成提供支持。