Cloning of the NM1killer toxin resistant gene in Saccharomyces cervisiae

酿酒酵母 NM1 杀手毒素抗性基因的克隆

• 批准号: 06660088
• 负责人: NAKAJIMA Tasuku
• 金额: $ 1.28万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06660088/
• 关键词: killer toxin; yeast cell wall; killer toxin resistant gene; beta-1,3-glucan; ベータ1, 3グルカン; ベータ1,3グルカン; 酵母の遺伝子

HM-1killer toxin secreted from Hansenula mrakii inhibits the growth of Saccharomyces cerevisiae cells by interfering with beta-1,3-glucan synthesis. A gene whose overexpression can endow Saccharomyces cerevisiae cells with resistant to HM-1 killer toxin was cloned. The gene, designated HKR1 contains 5.4-kb open reading frame. The HKR1 encodes a serine-and threonine-rich type 1 membrane protein with calcium-binding consensus sequence in the cytoplasmic domein. The null mutation of HKR1 is lethal and overexpression of HKR1 increases beta-1,3-glucan contents in cell wall. The partial disruption of HKR1 significantly reduced the level of beta-1,3-glucan synthase activities as well as cell wall beta-1,3-glucan contents, and altered the axial budding pattern of haploid cells. Immunofluorescednce microscopy with an antibody raised against Hkr1 expreseed in Escherichia coli revealed that HKR1 was predominantly localized on the cell surface. These results demonstrate that HKR1 encodes a cell surfase protein that regulates both cell wall beta-1,3-glucan synthesis and budding pattern.

由汉逊酵母分泌的HM-1杀手毒素通过干扰β-1，3-葡聚糖的合成来抑制酿酒酵母细胞的生长。克隆了一个能使酿酒酵母细胞对HM-1杀手毒素产生抗性的基因。该基因命名为HKR1，含有5.4kb的开放阅读框。HKR1编码一种富含丝氨酸和苏氨酸的1型膜蛋白，在细胞质Domein中具有与钙结合的共同序列。HKR1的零突变是致命的，过度表达会增加细胞壁中β-1，3-葡聚糖的含量。HKR1的部分破坏显著降低了β-1，3-葡聚糖合成酶活性和细胞壁β-1，3-葡聚糖含量，并改变了单倍体细胞的轴向萌发模式。免疫荧光显微镜观察发现，在大肠杆菌中表达的Hkr1主要定位于细胞表面。这些结果表明，HKR1编码一种细胞表面蛋白，调节细胞壁β-1，3-葡聚糖的合成和萌发模式。

项目成果

T. Yabeら: "HKR1 encodes a cell surface protein that both cell wall β-glucan synthesis and budding pattern in the yeast saccharomyces cerev, Tiae" J. Bacteriol.178. 477-483 (1996)

T. Yabe 等人：“HKR1 编码一种细胞表面蛋白，该蛋白参与酿酒酵母 (Tiae) 细胞壁 β-葡聚糖的合成和出芽模式”J. Bacteriol.177-483 (1996)。

T. Nakajimaら: "Chemical characterization of the galactomannan moiety in the cell wallgly copyoyteins of Aspergillus oryzae" J. Ferment. Bioeng.78. 472-476 (1994)

T. Nakajima 等人：“米曲霉细胞壁甘氨酸拷贝蛋白中半乳甘露聚糖部分的化学特征”J. Ferment. 472-476 (1994)。

S. Kasahara ら: "Involvement of cell wall β-Glucan in the action of HM-1 kiher to xin" FEBS Letters. 348. 27-32 (1994)

S. Kasahara 等人：“细胞壁 β-葡聚糖参与 HM-1 kiher to xin 的作用” FEBS Letters 348. 27-32 (1994)。

S. Kasaharaら: "Involvement of cell wall β-glucan in the action of HM-1 killer toxin." FEBS Letters. 348. 27-32 (1994)

S. Kasahara 等人：“细胞壁 β-葡聚糖参与 HM-1 杀伤毒素的作用。” FEBS Letters 348. 27-32 (1994)。

S.Kasahara, S.Inoue, T.Mio, T.Yamada, T.Nakajima, E.Ichishima, Y.Furuichi and H.Yamada: "Involvement of cell wall b-glucan in the action of HM-1 killer toxin." FEBS Letters. 348. 27-32 (1994)

S.Kasahara、S.Inoue、T.Mio、T.Yamada、T.Nakajima、E.Ichishima、Y.Furuichi 和 H.Yamada：“细胞壁 b-葡聚糖参与 HM-1 杀伤毒素的作用。