Molecular evolution of Aphanomyces cochlioides flavone receptor protein which functions in host recognition of the oomycetes zoospores.

丝丝菌黄酮受体蛋白的分子进化，其在卵菌游动孢子的宿主识别中发挥作用。

• 批准号: 24658103
• 负责人: HASHIDOKO Yasuyuki
• 金额: $ 2.58万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-24658103/
• 关键词: フラボン受容体; Aphanomyces cochlioides; VDAC; バレル構造

Aphanomyces cohlioides is a phytopathogen that attacks beet and spinach, using their characteristic secondary metabolite5-hydroxy-6,7-methylenedioxyflavone (cochliophilin A) as host recognition signal compound. We searched receptor protein to cause strong attractant activity of the B-ring non-substituted simple flavone. By using affinity column chromatography of solubilized membrane-binding proteins obtained from A. chochlioides zoospores, we successfully purified one candidate of the flavonoid receptor protein. Using a degenerate primer designed from the amino-acid sequence at the N-terminal of this protein, EST region of the target protein was successfully characterized to be aβ-barrel-type transmembrane protein of a 32 kDa, and named Aphanomyces cochliophilin A receptor protein 1 (ACCR1). From the mRNA library, full length of its cDNA was obtained. Amino acid sequence of ACCR1 had a high homology with voltage-dependent anion channel protein (VDAC), but ACCR1 ismissing 70AA from the C-terminus of VDAC. Our immunofluorescence assay for ACCR1 revealed that ACCR1 locates only cytoplasmic membrane but not on mitochondrion. All together, we proposed a hypothesis on molecular evolution of ACCR1 in Aphanomyces cochlioides.

类旋孢霉（Aphanomycescohlioides）是一种以甜菜和菠菜特有的次生代谢产物5-羟基-6，7-亚甲二氧基黄酮（旋孢菌素A）为寄主识别信号物质的植物病原菌。寻找引起B环非取代单黄酮强引诱活性的受体蛋白。用亲和柱层析法对A.从chochlioides游动孢子中分离纯化了一个类黄酮受体蛋白。根据该蛋白N端的氨基酸序列设计简并引物，成功鉴定了目的蛋白的EST区，其大小为32 kDa，为β-桶型跨膜蛋白，命名为Aphanomyces cochliophilin A receptor protein 1（ACCR 1）。从mRNA文库中获得了其全长cDNA。ACCR 1的氨基酸序列与电压依赖性阴离子通道蛋白（VDAC）有很高的同源性，但ACCR 1的C端缺失70个氨基酸。我们的免疫荧光检测显示ACCR 1只定位于细胞质膜而不定位于细胞膜上。综上所述，我们提出了一个假设，ACCR 1的分子进化的丝孢酵母cochlioides。