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构巢曲霉V类和VI类几丁质合成酶CsmA和CsmB,各自都具有一个类肌球蛋白运动结构域,它们执行着对菌丝顶端生长至关重要的补偿功能。

Aspergillus nidulans class V and VI chitin synthases CsmA and CsmB, each with a myosin motor-like domain, perform compensatory functions that are essential for hyphal tip growth.

作者信息

Takeshita Norio, Yamashita Shuichi, Ohta Akinori, Horiuchi Hiroyuki

机构信息

Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Tokyo 113-8657, Japan.

出版信息

Mol Microbiol. 2006 Mar;59(5):1380-94. doi: 10.1111/j.1365-2958.2006.05030.x.

Abstract

The polarized synthesis of cell wall components such as chitin is essential for the hyphal tip growth of filamentous fungi. The actin cytoskeleton is known to play important roles in the determination of hyphal polarity in Aspergillus nidulans. Previously, we suggested that CsmA, a chitin synthase with a myosin motor-like domain (MMD), was involved in polarized chitin synthesis in a manner dependent on the interaction between the MMD and the actin cytoskeleton. The genome database indicates that A. nidulans possesses another gene encoding another chitin synthase with an MMD. In this study, we characterized this gene, which we designated csmB. The csmB null mutants examined were viable, although they exhibited defective phenotypes, including the formation of balloons and intrahyphal hyphae and the lysis of subapical regions, which were similar to those obtained with csmA null mutants. Moreover, csmA csmB double null mutants were not viable. Mutants in which csmB was deleted and the expression of csmA was under the control of the alcA promoter were viable but severely impaired in terms of hyphal growth under alcA-repressing conditions. We revealed that CsmB with three copies of a FLAG epitope tag localized at the hyphal tips and forming septa, and that the MMD of CsmB was able to bind to actin filaments in vitro. These results suggest that CsmA and CsmB perform compensatory functions that are essential for hyphal tip growth.

摘要

几丁质等细胞壁成分的极性合成对于丝状真菌的菌丝顶端生长至关重要。已知肌动蛋白细胞骨架在构巢曲霉的菌丝极性确定中发挥重要作用。此前,我们提出CsmA,一种具有肌球蛋白运动样结构域(MMD)的几丁质合酶,以依赖于MMD与肌动蛋白细胞骨架之间相互作用的方式参与极性几丁质合成。基因组数据库表明构巢曲霉拥有另一个编码另一种具有MMD的几丁质合酶的基因。在本研究中,我们对这个我们命名为csmB的基因进行了表征。所检测的csmB缺失突变体是存活的,尽管它们表现出缺陷表型,包括气球状结构和菌丝内菌丝的形成以及亚顶端区域的裂解,这些与csmA缺失突变体所获得的表型相似。此外,csmA csmB双缺失突变体无法存活。csmB缺失且csmA的表达受alcA启动子控制的突变体是存活的,但在alcA抑制条件下菌丝生长严重受损。我们发现带有三个FLAG表位标签拷贝的CsmB定位于菌丝顶端并形成隔膜,并且CsmB的MMD在体外能够结合肌动蛋白丝。这些结果表明CsmA和CsmB执行对菌丝顶端生长至关重要的补偿功能。

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