Horiuchi H, Fujiwara M, Yamashita S, Ohta A, Takagi M
Department of Biotechnology, The University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan.
J Bacteriol. 1999 Jun;181(12):3721-9. doi: 10.1128/JB.181.12.3721-3729.1999.
We have found that the Aspergillus nidulans csmA gene encodes a novel protein which consists of an N-terminal myosin motor-like domain and a C-terminal chitin synthase domain (M. Fujiwara, H. Horiuchi, A. Ohta, and M. Takagi, Biochem. Biophys. Res. Commun. 236:75-78, 1997). To clarify the roles of csmA in fungal morphogenesis, we constructed csmA null mutants. The growth rate of the mutant colonies was almost the same as that of the wild-type strain, but hyphal growth was severely inhibited when a chitin-binding reagent, Calcofluor white or Congo red, was added to the medium. Moreover, morphological abnormalities in tip growth and septum formation were identified microscopically. Proliferation of intracellular new hyphae, called intrahyphal hyphae, which behaved as intrinsic hyphae, was the most striking phenotypic feature among them. These phenotypes were not suppressed when the only chitin synthase domain of csmA was expressed under the control of the alcA promoter, whereas they were suppressed when the intact form of csmA was expressed. Therefore, it was concluded that the product of csmA (CsmA) has important roles in polarized cell wall synthesis and maintenance of cell wall integrity and that the myosin motor-like domain is indispensable for these functions.
我们发现,构巢曲霉csmA基因编码一种新型蛋白质,该蛋白质由一个N端肌球蛋白运动样结构域和一个C端几丁质合酶结构域组成(M. Fujiwara、H. Horiuchi、A. Ohta和M. Takagi,《生物化学与生物物理研究通讯》236:75 - 78,1997年)。为了阐明csmA在真菌形态发生中的作用,我们构建了csmA缺失突变体。突变体菌落的生长速率与野生型菌株几乎相同,但当向培养基中添加几丁质结合试剂荧光增白剂或刚果红时,菌丝生长受到严重抑制。此外,在显微镜下还发现了顶端生长和隔膜形成的形态异常。细胞内新菌丝(称为菌丝内菌丝)的增殖最为显著,这些菌丝表现为固有菌丝。当csmA的唯一几丁质合酶结构域在alcA启动子的控制下表达时,这些表型并未得到抑制,而当完整形式的csmA表达时,它们则受到抑制。因此,得出结论,csmA的产物(CsmA)在极化细胞壁合成和维持细胞壁完整性方面具有重要作用,并且肌球蛋白运动样结构域对于这些功能是不可或缺的。