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The Septin AspB in Aspergillus nidulans Forms Bars and Filaments and Plays Roles in Growth Emergence and Conidiation

机译:构巢曲霉中的Septin AspB形成条和细丝并在生长萌发和分生过程中发挥作用

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摘要

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.
机译:在酵母中,Septins在母芽颈上形成环并充当扩散屏障。在动物中,Septins形成可以与其他细胞骨架成分共定位的细丝。在丝状真菌构巢曲霉中有五个septin基因,即aspA(酿酒酵母CDC11的直系同源物),aspB(酿酒酵母CDC3的直系同源物),aspC(酿酒酵母CDC12的直系同源物),aspD(S。的直系同源物)酿酒酵母CDC10)和aspE(仅在丝状真菌中发现)。以前有报道称aspB基因是表达最高的构巢曲霉Septin,并且是必不可少的。使用改良的基因靶向技术,我们发现删除aspB并不是致命的,但会导致分隔延迟,胚管和分支的出现增加,并且极大地减少了分生孢子。我们还发现,AspB绿色荧光蛋白(GFP)定位为分生孢子的隔片,分支和新兴层的环和项圈,以及分生孢子和菌丝中的条和细丝。在休眠和各向同性扩展的分生孢子中以及在菌丝的根尖下非生长区域中发现条形,并显示快速运动。随着胚芽管的出现,细丝形成,定位于菌丝和分支尖端,并显示出较慢的运动。所有可见的AspB-GFP结构均保留在ΔaspD中,并在ΔaspA和ΔaspC菌株中丢失。有趣的是,在ΔaspE突变体中,AspB-GFP的环,条和细丝在早期生长中可见,而AspB-GFP的条和细丝在隔垫形成后消失。 AspE直系同源物仅在丝状真菌中发现,表明这类Septins可能是高极性细胞中Septin条和细丝稳定性的必需条件。

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