Kakizaki Tetsuya, Sato Koh, Osakabe Keishi, Muraguchi Hajime
a Department of Biotechnology, Faculty of Bioresource Sciences, Akita Prefectural University.
b Faculty of Bioscience and Bioindustry, Tokushima University.
Mycoscience. 2024 Nov 20;65(6):307-316. doi: 10.47371/mycosci.2024.10.005. eCollection 2025.
The septins are observed at the hyphal tip and septum in the growing vegetative hyphae. In , septin dynamics have been suggested to be regulated by septin phosphorylation through protein kinases such as Cla4. To gain insight into the relationship between septins and Cla4 protein kinase (Cc.Cla4), we conducted disruption of the gene () by CRISPR/Cas9 system and analyzed the phenotypes of Δ strains. The vegetative hyphae of the Δ strains grow slowly in the zigzag shape and are highly branched compared to the wild-type hyphae. The enhanced green fluorescent protein (EGFP) tagged Cc.Cdc3 septin in the Δ strain was observed in the growing vegetative hyphae. Although EGFP-Cc.Cdc3 signals localized at the plasma membrane of the hyphal tip region, they did not accumulate as a dome with a hole at the hyphal tip as observed in the wild type. Instead, the Cc.Cdc3 was found to stay at the hyphal tip, which split into two tips thereafter. These results suggest that Cc.Cla4 is required for assembly of the hyphal tip apparatus that enables the rapid linear growth of the apical cells.
在生长的营养菌丝中，隔膜蛋白在菌丝顶端和隔膜处可见。在[具体研究中]，有研究表明隔膜蛋白的动态变化受Cla4等蛋白激酶介导的隔膜蛋白磷酸化调控。为深入了解隔膜蛋白与Cla4蛋白激酶（Cc.Cla4）之间的关系，我们利用CRISPR/Cas9系统对[基因名称]基因进行了敲除，并分析了Δ菌株的表型。与野生型菌丝相比，Δ菌株的营养菌丝呈锯齿状缓慢生长且分支较多。在Δ菌株中观察到增强型绿色荧光蛋白（EGFP）标记的Cc.Cdc3隔膜蛋白存在于生长的营养菌丝中。虽然EGFP-Cc.Cdc3信号定位于菌丝顶端区域的质膜，但它们并未像在野生型中那样在菌丝顶端积累形成有孔的穹顶状结构。相反，发现Cc.Cdc3停留在菌丝顶端，随后该顶端分裂为两个顶端。这些结果表明，Cc.Cla4是菌丝顶端装置组装所必需的，而该装置能使顶端细胞实现快速线性生长。
