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首页> 外文期刊>Molecular and Cellular Biology >nit-4, a pathway-specific regulatory gene of Neurospora crassa, encodes a protein with a putative binuclear zinc DNA-binding domain.
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nit-4, a pathway-specific regulatory gene of Neurospora crassa, encodes a protein with a putative binuclear zinc DNA-binding domain.

机译：nit-4是crus Neurospora crassa的一种通路特异性调控基因，它编码一种具有假定的双核锌DNA结合结构域的蛋白质。

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摘要

nit-4, a pathway-specific regulatory gene in the nitrogen circuit of Neurospora crassa, is required for the expression of nit-3 and nit-6, the structural genes which encode nitrate and nitrite reductase, respectively. The complete nucleotide sequence of the nit-4 gene has been determined. The predicted NIT4 protein contains 1,090 amino acids and appears to possess a single Zn(II)2Cys6 binuclear-type zinc finger, which may mediate DNA binding. Site-directed mutagenesis studies demonstrated that cysteine and other conserved amino acid residues in this possible DNA-binding domain are necessary for nit-4 function. A stretch of 27 glutamines, encoded by a CAGCAA repeating sequence, occurs in the C terminus of the NIT4 protein, and a second glutamine-rich domain occurs further upstream. A NIT4 protein deleted for the polyglutamine region was still functional in vivo. However, nit-4 function was abolished when both the polyglutamine region and the glutamine-rich domain were deleted, suggesting that the glutamine-rich domain might function in transcriptional activation. The homologous regulatory gene from Aspergillus nidulans, nirA, encodes a protein whose amino-terminal half has approximately 60% amino acid identity with NIT4 but whose carboxy terminus is completely different. A hybrid nit-4-nirA gene was constructed and found to function in N. crassa.
机译：nit-4是crus Neurospora crassa氮回路中的一种途径特异性调节基因，对于nit-3和nit-6的表达是必需的，nit-3和nit-6分别是编码硝酸盐和亚硝酸盐还原酶的结构基因。 nit-4基因的完整核苷酸序列已确定。预测的NIT4蛋白包含1,090个氨基酸，并且似乎拥有单个Zn（II）2Cys6双核型锌指，该锌指可能介导DNA结合。定点诱变研究表明，这种可能的DNA结合结构域中的半胱氨酸和其他保守氨基酸残基对于nit-4功能是必需的。由CAGCAA重复序列编码的27个谷氨酰胺片段出现在NIT4蛋白的C末端，第二个富谷氨酰胺结构域出现在更上游。缺失了聚谷氨酰胺区域的NIT4蛋白在体内仍然起作用。但是，当同时删除聚谷氨酰胺区域和富谷氨酰胺的结构域时，nit-4功能被取消，这表明富谷氨酰胺的域可能在转录激活中起作用。来自构巢曲霉的同源调控基因，nirA，编码一种蛋白质，其氨基末端一半与NIT4具有大约60％的氨基酸同一性，但其羧基末端却完全不同。构建了杂种nit-4-nirA基因，并发现其在猪笼草中起作用。

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