Bul1, a new protein that binds to the Rsp5 ubiquitin ligase in Saccharomyces cerevisiae
Bul1, a new protein that binds to the Rsp5 ubiquitin ligase in Saccharomyces cerevisiae
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Bul1，一种与酿酒酵母中的 Rsp5 泛素连接酶结合的新蛋白
DOI：
--
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发表时间：
1996
影响因子：
5.3
通讯作者：
Y. Kikuchi
中科院分区：
文献类型：
--
作者：
Hideki Yashiroda;T. Oguchi;Yuko Yasuda;A. Toh;Y. Kikuchi
我们鉴定了酿酒酵母的温度敏感突变体，其中微型染色体在高温下不稳定，并克隆了编码碱性亲水蛋白（110 kDa）的新基因。该基因的破坏导致了与原始突变相同的温度敏感生长。通过使用双杂交系统，我们进一步分离了编码hect（与E6-AP C末端同源）结构域的RSP5（反转Spt-表型）作为编码泛素连接酶的基因。因此，我们将基因命名为 BUL1（一种与泛素连接酶结合的蛋白质）。 BUL1 似乎参与了泛素化途径，因为编码泛素的高剂量 UBI1 部分抑制了 bul1 破坏体以及 rsp5 突变体的温度敏感性。 RSP5 和 BUL1 在多拷贝质粒上的共表达对于野生型细胞的有丝分裂生长具有毒性。脉冲追踪实验表明，野生型细胞中的Bul1保持稳定，而rsp5细胞中的Bul1条带几乎检测不到。由于通过免疫印迹分析确定两种菌株的蛋白质稳态水平相同，因此在没有完整 Rsp5 的情况下，Bul1 在免疫沉淀过程中可能很容易被降解。此外，Bul1 和 Rsp5 似乎都与通过蔗糖梯度离心分离的大复合物相关，并且 Rsp5 与 Bul1 共免疫沉淀。我们讨论了 Bul1 与 Rsp5 在蛋白质泛素化中共同发挥作用的可能性。
We characterized a temperature-sensitive mutant of Saccharomyces cerevisiae in which a mini-chromosome was unstable at a high temperature and cloned a new gene which encodes a basic and hydrophilic protein (110 kDa). The disruption of this gene caused the same temperature-sensitive growth as the original mutation. By using the two-hybrid system, we further isolated RSP5 (reverses Spt- phenotype), which encodes a hect (homologous to E6-AP C terminus) domain, as a gene encoding a ubiquitin ligase. Thus, we named our gene BUL1 (for a protein that binds to the ubiquitin ligase). BUL1 seems to be involved in the ubiquitination pathway, since a high dose of UBI1, encoding a ubiquitin, partially suppressed the temperature sensitivity of the bul1 disruptant as well as that of a rsp5 mutant. Coexpression of RSP5 and BUL1 on a multicopy plasmid was toxic for mitotic growth of the wild-type cells. Pulse-chase experiments revealed that Bul1 in the wild-type cells remained stable, while the bands of Bul1 in the rsp5 cells were hardly detected. Since the steady-state levels of the protein were the same in the two strains as determined by immunoblotting analysis, Bul1 might be easily degraded during immunoprecipitation in the absence of intact Rsp5. Furthermore, both Bul1 and Rsp5 appeared to be associated with large complexes which were separated through a sucrose gradient centrifugation, and Rsp5 was coimmunoprecipitated with Bul1. We discuss the possibility that Bul1 functions together with Rsp5 in protein ubiquitination.
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DOI：
--
发表时间：
1983-07
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
A. Hershko;H. Heller;S. Elias;A. Ciechanover
通讯作者：
A. Hershko;H. Heller;S. Elias;A. Ciechanover
突变 tRNA 影响酿酒酵母中 δ 介导的转录。
影响因子：
3.3
作者：
Happel,AM;Winston,F
通讯作者：
Winston,F
DOI：
10.1073/pnas.88.21.9578
发表时间：
1991-11-01
影响因子：
11.1
作者：
CHIEN, CT;BARTEL, PL;FIELDS, S
通讯作者：
FIELDS, S
影响因子：
10.5
作者：
EISENMANN, DM;ARNDT, KM;WINSTON, F
通讯作者：
WINSTON, F
影响因子：
7.5
作者：
HOCHSTRASSER, M
通讯作者：
HOCHSTRASSER, M