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binxman

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A HEX-1 crystal lattice required for Woronin body function in Neurospora crassa.

The Woronin body is a dense-core vesicle specific to filamentous ascomycetes (Euascomycetes), where it functions to seal the septal pore in response to cellular damage. The HEX-1 protein self-assembles to form this solid core of the vesicle. Here, we solve the crystal structure of HEX-1 at 1.8 A, which provides the structural basis of its self-assembly. The structure reveals the existence of three intermolecular interfaces that promote the formation of a three-dimensional protein lattice. Consistent with these data, self-assembly is disrupted by mutations in intermolecular contact residues and expression of an assembly-defective HEX-1 mutant results in the production of aberrantWoronin bodies, which possess a soluble noncrystalline core. This mutant also fails to complement a hex-1 deletion in Neurospora crassa, demonstrating that the HEX-1protein lattice is required for Woronin body function. Although both the sequence and the tertiary structure of HEX-1 are similar to those of eukaryotic initiation factor 5A (eIF-5A), the amino acids required for HEX-1 self-assembly and peroxisomal targeting are absent in eIF-5A. Thus, we propose that a new function has evolved following duplication of an ancestral eIF-5A gene and that this may define an important step in fungal evolution.
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fuaixiang

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binxman(金币+80, 翻译EPI+1): 谢谢!太感谢了!救命恩人啊 2011-10-21 23:09:45
粗糙链孢菌中乌头类生物碱功能所需的HEX-1晶格

乌头类生物碱是一种有致密核的囊泡,对丝状子囊菌(真子囊菌纲)有特效,其功能是封闭中隔孔以应对细胞损伤。HEX-1蛋白自组装而形成囊泡的实心核。在这里,我们在1.8 A处解开HEX-1的晶体结构,1.8 A提供了其自组装的结构基础。这种结构显示,有三个分子间接口促进三维蛋白晶格的形成。与这些数据一致的是,分子间接触残基的突变干扰了自组装,自组装缺陷HEX-1突变体的表达导致异常乌头类生物碱(具有水溶性非晶核)的产生。这种突变体也不能补足粗糙链孢菌中的hex-1缺失,证明HEX-1蛋白晶格是乌头类生物碱功能所必需的。尽管HEX-1的序列和三级结构都与真核起始因子5A(eIF-5A)类似,但是eIF-5A中缺少HEX-1自组装和过氧化物酶体定位所需的氨基酸。因此,我们认为,原始eIF-5A基因复制后形成了一种新功能,这可能决定了真菌进化中的一个重要步骤。
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