Hernández-Rodríguez Yainitza, Hastings Susan, Momany Michelle
University of Georgia, Athens, GA, USA.
Eukaryot Cell. 2012 Mar;11(3):311-23. doi: 10.1128/EC.05164-11. Epub 2012 Jan 13.
In yeast, septins form rings at the mother-bud neck and function as diffusion barriers. In animals, septins form filaments that can colocalize with other cytoskeletal elements. In the filamentous fungus Aspergillus nidulans there are five septin genes, aspA (an ortholog of Saccharomyces cerevisiae CDC11), aspB (an ortholog of S. cerevisiae CDC3), aspC (an ortholog of S. cerevisiae CDC12), aspD (an ortholog of S. cerevisiae CDC10), and aspE (found only in filamentous fungi). The aspB gene was previously reported to be the most highly expressed Aspergillus nidulans septin and to be essential. Using improved gene targeting techniques, we found that deletion of aspB is not lethal but results in delayed septation, increased emergence of germ tubes and branches, and greatly reduced conidiation. We also found that AspB-green fluorescent protein (GFP) localizes as rings and collars at septa, branches, and emerging layers of the conidiophore and as bars and filaments in conidia and hyphae. Bars are found in dormant and isotropically expanding conidia and in subapical nongrowing regions of hyphae and display fast movements. Filaments form as the germ tube emerges, localize to hyphal and branch tips, and display slower movements. All visible AspB-GFP structures are retained in ΔaspD and lost in ΔaspA and ΔaspC strains. Interestingly, in the ΔaspE mutant, AspB-GFP rings, bars, and filaments are visible in early growth, but AspB-GFP rods and filaments disappear after septum formation. AspE orthologs are only found in filamentous fungi, suggesting that this class of septins might be required for stability of septin bars and filaments in highly polar cells.
在酵母中,隔膜蛋白在母细胞-芽颈处形成环,并作为扩散屏障发挥作用。在动物中,隔膜蛋白形成细丝,可与其他细胞骨架成分共定位。在丝状真菌构巢曲霉中,有五个隔膜蛋白基因,即aspA(酿酒酵母CDC11的直系同源基因)、aspB(酿酒酵母CDC3的直系同源基因)、aspC(酿酒酵母CDC12的直系同源基因)、aspD(酿酒酵母CDC10的直系同源基因)和aspE(仅在丝状真菌中发现)。先前有报道称,aspB基因是构巢曲霉中表达量最高的隔膜蛋白基因,且是必需的。使用改进的基因靶向技术,我们发现缺失aspB并非致命,但会导致隔膜形成延迟、芽管和分支出现增加以及分生孢子形成大幅减少。我们还发现,AspB-绿色荧光蛋白(GFP)在隔膜、分支和分生孢子梗的新生层处定位为环和领圈,在分生孢子和菌丝中定位为条和细丝。条出现在休眠和各向同性扩展的分生孢子以及菌丝的亚顶端非生长区域,并表现出快速移动。细丝在芽管出现时形成,定位于菌丝和分支顶端,并表现出较慢的移动。所有可见的AspB-GFP结构在ΔaspD菌株中保留,而在ΔaspA和ΔaspC菌株中消失。有趣的是,在ΔaspE突变体中,AspB-GFP环、条和细丝在早期生长时可见,但在隔膜形成后,AspB-GFP杆和细丝消失。AspE直系同源基因仅在丝状真菌中发现,这表明这类隔膜蛋白可能是高极性细胞中隔膜蛋白条和细丝稳定性所必需的。