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粗糙脉孢菌中负责切割和ovc表型的基因编码一种卤代酸脱卤酶家族的蛋白质。

The Neurospora crassa gene responsible for the cut and ovc phenotypes encodes a protein of the haloacid dehalogenase family.

作者信息

Youssar Loubna, Schmidhauser Thomas J, Avalos Javier

机构信息

Departamento de Genética, Faculded de Biología, Universidad de Sevilla, Seville Spain.

出版信息

Mol Microbiol. 2005 Feb;55(3):828-38. doi: 10.1111/j.1365-2958.2004.04424.x.

Abstract

Light stimulation of carotenogenesis in Neurospora crassa, mediated by the White Collar proteins, is enhanced in some regulatory mutants, such as vivid and ovc. The gene responsible for the vivid mutation has been identified, but not the one responsible for the ovc phenotype. The ovc mutant is sensitive to high osmotic conditions and allelic with another mutant, cut, also osmosensitive but not affected in carotenogenesis. A phenotypic characterization of both strains is presented. Light induction of mRNA levels of the carotenoid genes al-1 and al-2, the regulatory gene wc-1 or the conidiation-specific gene con-10 is not significantly changed in the ovc mutant when compared with the wild type. We have identified the gene affected in the ovc mutant by complementation of osmosensitivity with a cosmid library. This gene, which we call cut-1, codes for an enzyme of the haloacid dehalogenase family, which includes different classes of phosphatases. cut-1 is able to restore the wild-type phenotype of the ovc and cut strains, confirming that they are affected in the same gene. DNA sequence analysis identified a point mutation in the cut mutant, leading to a truncated protein. The ovc mutant represents a deletion encompassing the entire gene and surrounding sequences. The cut-1 promoter contains putative regulatory elements involved in osmotic or thermal stress. We show that cut-1 transcription is low in illuminated or dark-grown cultures, and is induced by high osmotic conditions or by heat shock.

摘要

由白领蛋白介导的粗糙脉孢菌中类胡萝卜素生成的光刺激在一些调控突变体中增强,如生动突变体(vivid)和ovc突变体。导致生动突变的基因已被鉴定出来,但导致ovc表型的基因尚未确定。ovc突变体对高渗条件敏感,并且与另一个突变体cut等位,cut也对渗透压敏感,但在类胡萝卜素生成方面不受影响。本文展示了这两种菌株的表型特征。与野生型相比,在ovc突变体中,类胡萝卜素基因al-1和al-2、调控基因wc-1或分生孢子形成特异性基因con-10的mRNA水平的光诱导没有显著变化。我们通过用黏粒文库互补渗透压敏感性鉴定了ovc突变体中受影响的基因。这个我们称为cut-1的基因编码一种卤代酸脱卤酶家族的酶,该家族包括不同类别的磷酸酶。cut-1能够恢复ovc和cut菌株的野生型表型,证实它们在同一个基因上受到影响。DNA序列分析在cut突变体中鉴定出一个点突变,导致产生截短的蛋白质。ovc突变体代表一个包含整个基因及其周围序列的缺失。cut-1启动子含有参与渗透压或热应激的推定调控元件。我们表明,cut-1转录在光照或黑暗培养的菌株中较低,并由高渗条件或热休克诱导。

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