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Roles of NUDE and NUDF proteins of Aspergillus nidulans: Insights from intracellular localization and overexpression effects

机译:构巢曲霉的NUDE和NUDF蛋白的作用:细胞内定位和过表达效应的见解

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摘要

The NUDF protein of the filamentous fungus Aspergillus nidulans functions in the cytoplasmic dynein pathway. It binds several proteins, including the NUDE protein. Green fluorescent protein-tagged NUDF and NUDA (dynein heavy chain) localize to linearly moving dashes ("comets") that coincide with microtubule ends. Herein, deletion of the nudE gene did not eliminate the comets of NUDF and NUDA, but affected the behavior of NUDA. Comets were also observed with the green fluorescent protein-tagged NUDE and its nonfunctional C-terminal domain. In addition, overexpressed NUDA and NUDE accumulated in specks that were either immobile or bounced randomly. Neither comets nor specks were observed with the functional N-terminal domain of NUDE, indicating that these structures are not essential for NUDE function. Furthermore, NUDF overproduction totally suppressed deletion of the nudE gene. This implies that the function of NUDE is secondary to that of NUDF. Unexpectedly, NUDF overproduction inhibited one conditional nudA mutant and all tested apsA mutants. An allele-specific interaction between the nudF and nudA genes is consistent with a direct interaction between NUDF and dynein heavy chain. Because APSA and its yeast homolog Num1p are cortical proteins, an interaction between the nudF and apsA genes suggests a role for NUDF at the cell cortex. [References: 76]
机译:丝状真菌构巢曲霉的NUDF蛋白在细胞质动力蛋白途径中起作用。它结合几种蛋白质,包括NUDE蛋白质。带绿色荧光蛋白标签的NUDF和NUDA(动力蛋白重链)定位于与微管末端重合的线性移动虚线(“彗星”)。在此,缺失nudE基因并不能消除NUDF和NUDA的彗星,但是影响了NUDA的行为。还观察到带有绿色荧光蛋白标签的NUDE及其无功能的C末端结构域的彗星。此外,过表达的NUDA和NUDE聚集在固定不动或随机反弹的斑点中。 NUDE的功能性N末端结构域未观察到彗星或斑点,这表明这些结构对于NUDE功能不是必需的。此外,NUDF过度生产完全抑制了nudE基因的缺失。这意味着NUDE的功能仅次于NUDF。出乎意料的是,NUDF的过量生产抑制了一个条件性nudA突变体和所有测试的apsA突变体。 nudF和nudA基因之间的等位基因特异性相互作用与NUDF和动力蛋白重链之间的直接相互作用是一致的。由于APSA及其酵母同源物Num1p是皮质蛋白,因此nudF和apsA基因之间的相互作用提示NUDF在细胞皮质中的作用。 [参考:76]

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