Zhang Shizhu, Zheng Hailin, Long Nanbiao, Carbó Natalia, Chen Peiying, Aguilar Pablo S, Lu Ling
Jiangsu Key Laboratory for Microbes and Functional Genomics, Engineering and Technology Research Center for Microbiology, College of Life Sciences, Nanjing Normal University, Nanjing, Jiangsu, China.
Eukaryot Cell. 2014 Feb;13(2):295-303. doi: 10.1128/EC.00257-13. Epub 2013 Dec 27.
Calcium-mediated signaling pathways are widely employed in eukaryotes and are implicated in the regulation of diverse biological processes. In Saccharomyces cerevisiae, at least two different calcium uptake systems have been identified: the high-affinity calcium influx system (HACS) and the low-affinity calcium influx system (LACS). Compared to the HACS, the LACS in fungi is not well known. In this study, FigA, a homolog of the LACS member Fig1 from S. cerevisiae, was functionally characterized in the filamentous fungus Aspergillus nidulans. Loss of figA resulted in retardant hyphal growth and a sharp reduction of conidial production. Most importantly, FigA is essential for the homothallic mating (self-fertilization) process; further, FigA is required for heterothallic mating (outcrossing) in the absence of HACS midA. Interestingly, in a figA deletion mutant, adding extracellular Ca(2+) rescued the hyphal growth defects but could not restore asexual and sexual reproduction. Furthermore, quantitative PCR results revealed that figA deletion sharply decreased the expression of brlA and nsdD, which are known as key regulators during asexual and sexual development, respectively. In addition, green fluorescent protein (GFP) tagging at the C terminus of FigA (FigA::GFP) showed that FigA localized to the center of the septum in mature hyphal cells, to the location between vesicles and metulae, and between the junctions of metulae and phialides in conidiophores. Thus, our findings suggest that FigA, apart from being a member of a calcium uptake system in A. nidulans, may play multiple unexplored roles during hyphal growth and asexual and sexual development.
钙介导的信号通路在真核生物中广泛应用,参与多种生物过程的调控。在酿酒酵母中,已鉴定出至少两种不同的钙摄取系统:高亲和力钙内流系统(HACS)和低亲和力钙内流系统(LACS)。与HACS相比,真菌中的LACS尚不为人所知。在本研究中,丝状真菌构巢曲霉中对酿酒酵母LACS成员Fig1的同源物FigA进行了功能表征。FigA缺失导致菌丝生长迟缓,分生孢子产量急剧下降。最重要的是,FigA对于同宗配合(自体受精)过程至关重要;此外,在缺乏HACS成员MidA的情况下,异宗配合(杂交)也需要FigA。有趣的是,在FigA缺失突变体中,添加细胞外Ca(2+)可挽救菌丝生长缺陷,但无法恢复无性和有性繁殖。此外,定量PCR结果显示,FigA缺失显著降低了brlA和nsdD的表达,brlA和nsdD分别是无性和有性发育过程中的关键调节因子。此外,在FigA的C末端标记绿色荧光蛋白(FigA::GFP)表明,FigA定位于成熟菌丝细胞的隔膜中心、分生孢子梗中囊泡与梗基之间以及梗基与小梗连接处之间的位置。因此,我们的研究结果表明,FigA除了是构巢曲霉钙摄取系统的成员外,可能在菌丝生长以及无性和有性发育过程中发挥多种未被探索的作用。