Chung Kuang-Ren, Daub Margaret E, Kuchler Karl, Schüller Christoph
Citrus Research and Education Center and Department of Plant Pathology, IFAS, University of Florida, Lake Alfred, FL 33850, USA.
Biochem Biophys Res Commun. 2003 Mar 7;302(2):302-10. doi: 10.1016/s0006-291x(03)00171-2.
The Cercospora nicotianae CRG1 gene is involved in cellular resistance to the perylenequinone toxin, cercosporin, that generates highly toxic singlet oxygen upon exposure to light. The entire open reading frame (ORF) of CRG1 was isolated and sequenced. The gene contains an ORF of 1950bp including a 65-bp intron. The predicted 650 amino acid CRG1 protein contains a Cys(6)Zn(2) binuclear cluster DNA-binding motif with homology to various fungal regulatory proteins, indicating that CRG1 may act functionally as a transcription activator. Targeted gene disruption of CRG1 resulted in mutants that are partially sensitive to cercosporin and reduced in cercosporin production. Genetic complementation revealed that CRG1 fully restored cercosporin resistance, but only slightly restored cercosporin production in a UV-derived mutant (CS10) containing a single nucleotide substitution in crg1. Complementation of a crg1-null mutant, however, yielded strains that are similar to the wild-type in both phenotypes. These results indicate that the transcription regulator CRG1 is involved in the activation of genes associated with cercosporin resistance and production in the fungus Cercospora nicotianae.
烟草尾孢菌CRG1基因参与细胞对苝醌毒素——尾孢菌素的抗性,尾孢菌素在光照下会产生剧毒的单线态氧。分离并测序了CRG1的完整开放阅读框(ORF)。该基因包含一个1950bp的ORF,其中有一个65bp的内含子。预测的650个氨基酸的CRG1蛋白含有一个Cys(6)Zn(2)双核簇DNA结合基序,与各种真菌调节蛋白具有同源性,这表明CRG1可能在功能上作为转录激活因子发挥作用。对CRG1进行靶向基因敲除导致突变体对尾孢菌素部分敏感,且尾孢菌素产量降低。遗传互补分析表明,CRG1完全恢复了对尾孢菌素的抗性,但在crg1中含有单核苷酸取代的紫外线衍生突变体(CS10)中,仅略微恢复了尾孢菌素的产量。然而,对crg1缺失突变体的互补产生了在两种表型上都与野生型相似的菌株。这些结果表明,转录调节因子CRG1参与激活烟草尾孢菌中与尾孢菌素抗性和产生相关的基因。