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冠状蛋白是粗糙脉孢菌菌丝的内吞环的一个组成部分,对正常生长和形态发生是必需的。

Coronin is a component of the endocytic collar of hyphae of Neurospora crassa and is necessary for normal growth and morphogenesis.

机构信息

Departamento de Microbiología, Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE), Ensenada, Baja California, México.

出版信息

PLoS One. 2012;7(5):e38237. doi: 10.1371/journal.pone.0038237. Epub 2012 May 31.

Abstract

Coronin plays a major role in the organization and dynamics of actin in yeast. To investigate the role of coronin in a filamentous fungus (Neurospora crassa), we examined its subcellular localization using fluorescent proteins and the phenotypic consequences of coronin gene (crn-1) deletion in hyphal morphogenesis, Spitzenkörper behavior and endocytosis. Coronin-GFP was localized in patches, forming a subapical collar near the hyphal apex; significantly, it was absent from the apex. The subapical patches of coronin colocalized with fimbrin, Arp2/3 complex, and actin, altogether comprising the endocytic collar. Deletion of crn-1 resulted in reduced hyphal growth rates, distorted hyphal morphology, uneven wall thickness, and delayed establishment of polarity during germination; it also affected growth directionality and increased branching. The Spitzenkörper of Δcrn-1 mutant was unstable; it appeared and disappeared intermittently giving rise to periods of hyphoid-like and isotropic growth respectively. Uptake of FM4-64 in Δcrn-1 mutant indicated a partial disruption in endocytosis. These observations underscore coronin as an important component of F-actin remodeling in N. crassa. Although coronin is not essential in this fungus, its deletion influenced negatively the operation of the actin cytoskeleton involved in the orderly deployment of the apical growth apparatus, thus preventing normal hyphal growth and morphogenesis.

摘要

冠蛋白在酵母中肌动蛋白的组织和动力学中起主要作用。为了研究冠蛋白在丝状真菌(粗糙脉孢菌)中的作用,我们使用荧光蛋白研究了其亚细胞定位,以及冠蛋白基因(crn-1)缺失对菌丝形态发生、Spitzenkörper 行为和内吞作用的表型影响。冠蛋白-GFP 定位于斑点中,在菌丝顶端附近形成亚顶端环;值得注意的是,它不在顶端。冠蛋白的亚顶端斑点与 fimbrin、Arp2/3 复合物和肌动蛋白共定位,共同构成内吞环。crn-1 缺失导致菌丝生长速度降低、菌丝形态扭曲、壁厚度不均匀以及在萌发过程中极性建立延迟;它还影响生长方向性并增加分支。Δcrn-1 突变体的 Spitzenkörper 不稳定;它间歇性出现和消失,分别导致菌丝状和各向同性生长。在 Δcrn-1 突变体中,FM4-64 的摄取表明内吞作用部分受到破坏。这些观察结果强调了冠蛋白作为 N. crassa 中 F-actin 重塑的重要组成部分。尽管冠蛋白在这种真菌中不是必需的,但它的缺失会对参与顶端生长装置有序部署的肌动蛋白细胞骨架的运作产生负面影响,从而阻止正常的菌丝生长和形态发生。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b3cc/3365027/c20637c2a86b/pone.0038237.g001.jpg

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