Regulation of cellular functions by the ubiquitin-proteasome system in Saccharomyces cerevisiae

酿酒酵母泛素蛋白酶体系统对细胞功能的调节

• 批准号: 08454242
• 负责人: TOH-E Akio
• 金额: $ 5.06万
• 依托单位: Department of Biological Sciences, Graduate School of Science, University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08454242/
• 关键词: proteasome; ubiquitin; cell cycle; proteolysis; Cdk; yeast; ユビキチン; SUN1; SUN2

Nin1P,a component of the 26S proteasome of Saccharomyces cerevisiae, is required for activation of Cdc28p kinase at the G1/S and G2/M boundaries. By exploiting the temperature sensitive phenotype of the nin1-1 mutant, we have screened for genes encoding proteins with related functions to Nin1P and have cloned and characterized two new multicopy suppressors, SUN1 and SUN2, of the nin1-1 mutation. SUN1 can suppress a null nin1 mutation, whereas SUN2, an essential gene, does not. Sun1p is a 268 amino acid protein which shows strong similarity to MBP1 of Arabidopsis thaliana, a homolog of the S5a subunit of the human 26S proteasome. Sun1p binds ubiquitin-lysozyme conjugates as do S5a and MBP1. Sun2p (523 amino acids) was found to be homologous to the p58 subunit of the human 26S proteasome. cDNA encoding the p58 component was cloned. Furthermore, expression of a derivative of p58 from which the N-terminal 150 amino acids had been removed restored the function of a null allele of SUN2. During glycerol density gradient centrifugation, both Sun1p and Sun2p comigrated with the known proteasome components. These results, as well as, other structural and functional studies, insicate that both Sun1p and Sun2p are components of the regulatory module of the yeast 26S proteasome

Nin1P 是酿酒酵母 26S 蛋白酶体的一个组成部分，是在 G1/S 和 G2/M 边界激活 Cdc28p 激酶所必需的。通过利用nin1-1突变体的温度敏感表型，我们筛选了编码与Nin1P具有相关功能的蛋白质的基因，并克隆并表征了nin1-1突变体的两个新的多拷贝抑制子SUN1和SUN2。 SUN1 可以抑制 nin1 无效突变，而必需基因 SUN2 则不能。 Sun1p 是一种 268 个氨基酸的蛋白质，与拟南芥的 MBP1 具有很强的相似性，MBP1 是人类 26S 蛋白酶体 S5a 亚基的同源物。 Sun1p 与 S5a 和 MBP1 一样结合泛素-溶菌酶缀合物。 Sun2p（523 个氨基酸）被发现与人 26S 蛋白酶体的 p58 亚基同源。克隆了编码p58成分的cDNA。此外，N端150个氨基酸被去除的p58衍生物的表达恢复了SUN2无效等位基因的功能。在甘油密度梯度离心过程中，Sun1p 和 Sun2p 均与已知的蛋白酶体成分发生共迁移。这些结果以及其他结构和功能研究表明 Sun1p 和 Sun2p 都是酵母 26S 蛋白酶体调节模块的组成部分

项目成果

K.Kominami, N.Okura, M.Kawamura et al.: "Yeast counterparts of subunits S5a and p58(S3)of the human 26S Proteasome are encoded by two multicopy suppressors of nin1-1." Mol.Biol.Cell.8. 171-187 (1997)

K.Kominami、N.Okura、M.Kawamura 等人：“人类 26S 蛋白酶体的亚基 S5a 和 p58(S3) 的酵母对应物由 nin1-1 的两个多拷贝抑制子编码。”

Yoshida,M.,Oguchi,T.,Yasuda,Y.,Toh-e,A.,and Kikuch,Y.: "Bul1,a new binding protein of the Rsp5-ubiquitin Qigase in Sacchanomyces cerevisiae" Mol.Biol.Cell. 16. 3255-3263 (1996)

Yoshida,M.、Oguchi,T.、Yasuda,Y.、Toh-e,A. 和 Kikuch,Y.：“Bul1，酿酒酵母中 Rsp5-泛素 Qigase 的新结合蛋白”Mol.Biol.Cell

H.Yashiroda, T.Oguchi, Y.Yasuda, A.Toh-e and Y.Kikuchi: "Bull,a new protein that binds to the Rsp5 ubiquitin ligase in Saccharomyces cerevisiae." Mol.Cell.Biol.16. 3255-3263 (1996)

H.Yashiroda、T.Oguchi、Y.Yasuda、A.Toh-e 和 Y.Kikuchi：“Bull，一种与酿酒酵母中的 Rsp5 泛素连接酶结合的新蛋白质。”

M.Kawamura, K.Kominami, J.Takeuchi and A.Toh-e: "A multicopy suppressor of nin1-1 of the yeast Saccharomyces cerevisiae is a couterpart of the Drosophila melanogaster diphenol oxidase A2 gene,Dox-A2." Mol.Gen.Genet.251. 146-152 (1996)

M.Kawamura、K.Kominami、J.Takeuchi 和 A.Toh-e：“酿酒酵母 nin1-1 的多拷贝抑制因子是果蝇二酚氧化酶 A2 基因 Dox-A2 的配对基因。”

Andoh,T., Kato,T.Jr., Matsui,Y., Tuh-e,A: "Phosphoinositide,specil-c phospholirase C forms a complex with 14-3-3 priteins and is in volved inexpression of UV resistance in fission yeast" Mol.Gen.Genef.(in press).

Andoh,T.、Kato,T.Jr.、Matsui,Y.、Tuh-e,A：“磷酸肌醇、specil-c 磷酸酶 C 与 14-3-3 蛋白形成复合物，并参与抗紫外线的表达